Antibacterial agents

ABSTRACT

Antibacterial compounds of formula (I) are provided: 
     
       
         
         
             
             
         
       
     
     as well as stereoisomers, pharmaceutically acceptable salts, esters, and prodrugs thereof; pharmaceutical compositions comprising such compounds; methods of treating bacterial infections by the administration of such compounds; and processes for the preparation of such compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/223,971, filed Mar. 24, 2014 (allowed), which is a continuation ofU.S. patent application Ser. No. 12/635,551, filed Dec. 10, 2009 (nowabandoned), which is a continuation of International PCT Application No.PCT/US2008/066766, filed Jun. 12, 2008, which claims the benefit under35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 60/943,494,filed Jun. 12, 2007. The foregoing applications are incorporated hereinby reference in their entireties.

STATEMENT OF GOVERNMENT INTEREST

This invention was made with government support under Contract No.HDTRA1-07-C-0079 awarded by the United States Department of Defense. Thegovernment has certain rights in this invention.

BACKGROUND OF THE INVENTION

Field of the Invention

This invention pertains generally to treating infections cause bygram-negative bacteria. More specifically, the invention describedherein pertains to treating gram-negative infections by inhibitingactivity of UDP-3-O—(R-3-hydroxydecanoyl)-N-acetylglucosaminedeacetylase (LpxC). The present invention provides small moleculeinhibitors of LpxC, pharmaceutical formulations containing suchinhibitors, methods of treating patients with such pharmaceuticalformulations, and methods of preparing such pharmaceutical formulationsand inhibitors. The inhibitors can be used to treat gram-negativeinfections of patients alone and in combination with otherantibacterials.

Description of the Related Art

Over the past several decades, the frequency of antimicrobial resistanceand its association with serious infectious diseases have increased atalarming rates. The increasing prevalence of resistance among nosocomialpathogens is particularly disconcerting. Of the over 2 millionnosocomial infections occurring each year in the United States, 50 to60% are caused by antimicrobial-resistant strains of bacteria. This highrate of resistance increases the morbidity, mortality, and costsassociated with nosocomial infections. In the United States, nosocomialinfections are thought to contribute to or cause more than 77,000 deathsper year and cost approximately $5 to $10 billion annually. Amonggram-positive organisms, the most important resistant pathogens aremethicillin-(oxacillin-)resistant Staphylococcus aureus,β-lactam-resistant and multidrug-resistant pneumococci, andvancomycin-resistant enterococci. Important causes of gram-negativeresistance include extended-spectrum β-lactamases (ESBLs) in Klebsiellapneumoniae, Escherichia coli, and Proteus mirabilis, high-levelthird-generation cephalosporin (Amp C) β-lactamase resistance amongEnterobacter species and Citrobacter freundii, and multidrug-resistancegenes observed in Pseudomonas aeruginosa, Acinetobacter, andStenotrophomonas maltophilia (see Jones, R. N., “Resistance patternsamong nosocomial pathogens: Trends over the past few years” Chest.,2001, 119 (Supp 2), 397S-404S).

The problem of antibacterial resistance is compounded by the existenceof bacterial strains resistant to multiple antibacterials. For example,Pseudomonas aeruginosa isolates resistant to fluoroquinolones arevirtually all resistant to additional antibacterials (see Sahm, D. F. etal., “Evaluation of current activities of fluoroquinolones againstgram-negative bacilli using centralized in vitro testing and electronicsurveillance” Antimicrobial Agents and Chemotherapy, 2001, 45, 267-274).

Thus there is a need for new antibacterials, particularly antibacterialswith novel mechanisms of action. Most of the antibacterial discoveryeffort in the pharmaceutical industry is aimed at development of drugseffective against gram-positive bacteria. However, there is also a needfor new gram-negative antibacterials. Gram-negative bacteria are ingeneral more resistant to a large number of antibacterials andchemotherapeutic agents than are gram-positive bacteria. A survey ofrecently reported antibacterials of natural origin showed that over 90%lacked activity against Escherichia coli, although they were activeagainst gram-positive bacteria. The outer membrane of gram-negativebacteria contributes to this intrinsic resistance by acting as anefficient permeability barrier, because the narrow porin channels limitthe penetration of hydrophilic solutes and the low fluidity of thelipopolysaccharide leaflet slows down the inward diffusion of lipophilicsolutes. A second mechanism also contributes to the intrinsic resistanceof gram-negative bacteria. Recent studies showed that multiple drugefflux pumps, sometimes with unusually broad specificity, act as thissecond factor to create the general intrinsic resistance ofgram-negative bacteria. When their expression levels are elevated as aconsequence of physiological regulation or genetic alteration, they canfrequently produce impressive levels of resistance to a wide variety ofantimicrobial agents (see Nikaido H., “Antibacterial resistance causedby gram-negative multidrug efflux pumps” Clinical Infectious Diseases,1998, 27 (Supp 1), S32-41).

Historically, most development of antimicrobial agents has beenrelatively empirical. Active compounds have generally been found viascreening soil, sewage, water, and other natural substances to detectantimicrobial-producing organisms, or by screening various chemicalcompounds. Once a leading candidate has been found and its chemicalstructure determined, a series of analogs is made to identify an optimalcompound for further clinical development. A more rational approachinvolves the defining of new targets, such as genes or enzymaticfunctions, responsible for a crucial cellular essential activity. Oncethis has been done, inhibitors or blockers of the function or geneproduct can be developed.

In order to identify potential targets for novel gram-negativeantibacterial agents, studies aimed at identifying all essential andimportant genes in Pseudomonas aeruginosa have been performed. Among theessential genes identified was LpxC, that encodes the enzymeuridyldiphospho-3-O—(R-hydroxydecanoyl)-N-acetylglucosamine deacetylase(LpxC). This enzyme is the first committed step in the synthesis oflipid A, the lipid moiety of lipopolysaccharide, that is an essentialcomponent of all gram-negative bacteria. It therefore is an attractivetarget for novel antibacterials. In order to be useful as antibacterialagents, LpxC inhibitors would not only have to inhibit the enzymaticactivity of LpxC from a variety of bacteria, but would have to defeatthe intrinsic resistance mechanisms of gram-negative bacteria, asdescribed above (i.e., they would have to penetrate the outer membraneand be relatively unsusceptible to multidrug efflux pumps).

To date, researchers have identified a few compounds with antibacterialactivity that target lipid A biosynthesis. For example, InternationalPCT Publication No. WO 97/42179 to Patchett et al. discloses compoundsof the formula:

The compounds possess activity against certain gram-negative organisms,for example Escherichia coli, but are not active against other medicallyimportant gram-negative bacteria, for example Pseudomonas aeruginosa.Subsequent studies have found that the primary reason for theirinactivity against particular, medically important gram-negativebacteria is their poor ability to inhibit P. aeruginosa LpxC; efflux bythe major multidrug efflux pump or inability to penetrate the outermembrane were not the critical factors.

Jackman et al. (J. Biol. Chem., 2000, 275(15), 11002-11009) discuss themechanism of lipid A biosynthesis in the context of gram-negativebacteria and disclose a new class of hydroxamate-containing inhibitorsof LpxC. Wyckoff et al. (Trends in Microbiology, 1998, 6(4), 154-159)discuss the role of LpxC in lipid A biosynthesis and its role inregulation and disclose a few oxazoline hydroxamic acids that inhibitbacterial growth. However, Wyckoff et al. also discuss the shortcomingsof the available deacetylase inhibitors as bactericidal agents againstPseudomonas and note that more work is needed to be done in the area.

U.S. Patent Application Publication No. 2001/0053555 (published Dec. 20,2001, corresponding to International PCT Publication No. WO 98/18754published May 7, 1998) discloses a combinatorial library ofhydroxylamine, hydroxamic acid, hydroxyurea and hydroxylsulfonamidecompounds purported to be potentially useful as inhibitors ofmetalloproteases, and U.S. Pat. No. 6,281,245 claims a method ofinhibiting a deformylase enzyme by administering one of thehydroxylamine compounds from the combinatorial library disclosed in U.S.Patent Application Publication No. 2001/0053555. Related to theforegoing patent publications is International PCT Publication No. WO99/57097 (published Nov. 11, 1999) that discloses a method of solidphase synthesis of the hydroxylamine library of compounds.

International PCT Publication No. WO 00/61134 to British BiotechPharmaceuticals Limited (published Oct. 19, 2000) discloses compounds ofthe formula:

The compounds are useful as antimicrobial agents and are believed tohave bactericidal activity due, at least in part, to intracellularinhibition of bacterial polypeptide deformylase.

In earlier International PCT Publication No. WO 99/39704 to BritishBiotech Pharmaceuticals Limited (published Aug. 12, 1999), compounds ofthe following formula were disclosed:

The compounds are useful as antimicrobial agents useful againstgram-negative and gram positive bacteria.

De Novo Pharmaceuticals LTD disclosed in International PCT PublicationNo. WO 02/50081 (published Jun. 27, 2002), certain antibacterial andantiprotozoal agents having the formulae shown below:

The patent publication discusses that the antibacterial activity is due,at least in part, to intracellular inhibition of bacterial polypeptidedeformylase.

More recently, certain compounds having activity against gram-negativebacterial infections were disclosed in U.S. Patent ApplicationPublication No. 2004/0229955 (published Nov. 18, 2004).

Although there have been advances in the field, there remains a need forLpxC inhibitors that have activity as bactericidal agents againstgram-negative bacteria. It is, accordingly, an object of this inventionto provide compounds and combinations of such compounds for use in thepreparation of antibacterials and other pharmaceuticals capable ofinhibiting gram-negative bacterial infections.

BRIEF SUMMARY OF THE INVENTION

The present invention provides novel compounds, pharmaceuticalformulations including the compounds, methods of inhibitingUDP-3-O—(R-3-hydroxydecanoyl)-N-acetylglucosamine deacetylase (LpxC),and methods of treating gram-negative bacterial infections.

In one aspect, the present invention provides compounds having thefollowing formula (I):

including stereoisomers, pharmaceutically acceptable salts, esters, andprodrugs thereof.

In a first embodiment, the present invention provides compounds offormula (I) wherein:

E is selected from the group consisting of:

-   -   (1) H,    -   (2) substituted or unsubstituted C₁-C₆-alkyl,    -   (3) substituted or unsubstituted C₂-C₆-alkenyl,    -   (4) substituted or unsubstituted C₂-C₆-alkynyl,    -   (5) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (6) substituted or unsubstituted aryl,    -   (7) substituted or unsubstituted heterocyclyl, and    -   (8) substituted or unsubstituted heteroaryl;

L is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₁-C₆-alkyl,    -   (2) —(NR^(3L))₀₋₁—(CH₂)₀₋₄—NR^(3L)—(CH₂)₀₋₄—,    -   (3) —(NR^(3L))₀₋₁—C(R^(1L),R^(2L))—NR^(3L), —C(R^(1L),R^(2L))—,    -   (4) —C(R^(1L),R^(2L))—O—C(R^(1L),R^(2L))—,    -   (5) —(CH₂)₀₋₄—NR^(3L)—C(R^(1L),R^(2L))—CONH—(CH₂)₀₋₄—,    -   (6) —CO—C(R^(1L),R^(2L))—NHCO—,    -   (7) —CONR^(3L)—,    -   (8) —NR^(3L)CO—,    -   (9) —NR^(3L)—,    -   (10) —SO₂NR^(3L)—,    -   (11) —NR^(3L)—C(═O)—NR^(3L)—,    -   (12) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (13) substituted or unsubstituted aryl,    -   (14) substituted or unsubstituted heterocyclyl, and    -   (15) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1L), R^(2L), and R^(3L) is independently selected            from the group consisting of:            -   (a) H,            -   (b) substituted or unsubstituted C₁-C₆-alkyl,            -   (c) C₁-C₆-alkyl substituted with aryl,            -   (d) C₁-C₆-alkyl substituted with heterocyclyl, and            -   (e) C₁-C₆-alkyl substituted with heteroaryl,    -   or R^(1L) and R^(3L), together with the atoms to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 3 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S;

D is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

G is selected from the group consisting of:

-   -   (1) —NR^(1G)C(═O)—,    -   (2) —C(═O)NR^(1G)—,    -   (3) —(CH₂)₀₋₄NHCH₂C(═O)NR^(1G)—,    -   (4) —CR^(2G)═CR^(2G)—,    -   (5) —S(═O)—,    -   (6) —SO₂—,    -   (7) —C(R^(3G))₂—S(═O)—,    -   (8) —S(═O)—C(R^(3G))₂—,    -   (9) —C(R^(3G))₂—SO₂—,    -   (10) —SO₂—C(R^(3G))₂—,    -   (11) —CR^(3G)═CR^(3G)—CR^(3G)═CR^(3G)—,    -   (12) —C(R^(3G))₂—,    -   (13) —CR^(3G)═CR^(3G)—C≡C—,    -   (14) —C≡C—CR^(3G)═CR^(3G)—,    -   (15) —C(═O)—C≡C—,    -   (16) —C≡C—C(═O)—,    -   (17) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (18) substituted or unsubstituted aryl,    -   (19) substituted or unsubstituted heterocyclyl, and    -   (20) substituted or unsubstituted heteroaryl,    -   wherein:        -   R^(1G) is substituted or unsubstituted C₁-C₆-alkyl;        -   each R^(2G) is independently selected from the group            consisting of H, a halogen atom, and substituted or            unsubstituted C₁-C₆-alkyl, and at least one R^(2G) is not H;            and        -   R^(3G) is selected from the group consisting of H, a halogen            atom, and substituted or unsubstituted C₁-C₆-alkyl;

Y is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

X is selected from the group consisting of:

-   -   (1) —(C═O)NR₄—,    -   (2) —C₁-C₆-alkyl-(C═O)NR₄—,    -   (3) —C₂-C₆-alkenyl-(C═O)NR₄—,    -   (4) —C₂-C₆-alkynyl-(C═O)NR₄—,    -   (5) —CH₂NR₄—,    -   (6) —SO₂NR₄—,    -   (7) —S(═O)NR₄—,    -   (8) —NR₄C(═O)—, and    -   (9) —NR₄—,    -   or X and A, together with the atoms to which they are attached        can form a heterocyclic ring, having from 5 to 8 ring atoms,        wherein 1-2 ring atoms of the heterocyclic ring are selected        from N, O and S,    -   or when Y is a bicyclic substituted or unsubstituted        heterocyclyl or heteroaryl, then X is absent;

R₃ is H or substituted or unsubstituted C₁-C₆-alkyl, or R₃ and A,together with the atom to which they are attached can form a substitutedor unsubstituted 3-10 membered cycloalkyl or a heterocyclic ring, havingfrom 3 to 10 ring atoms, wherein 1-4 ring atoms of the heterocyclic ringare selected from N, O and S;

R₄ is (1) H or substituted or unsubstituted C₁-C₆-alkyl, or (2) R₄ andA, together with the atoms to which they are attached can form asubstituted or unsubstituted heterocyclic ring, having from 3 to 8 ringatoms, wherein 1-2 ring atoms of the heterocyclic ring are selected fromN, O and S, or (3) R₄ and Y, together with the atoms to which they areattached, form a bicyclic substituted or unsubstituted heterocyclyl orheteroaryl;

n is an integer from 0-6;

A is selected from the group consisting of:

-   -   (1) H,    -   (2) —(CH₂)₀₋₄C(R^(1a),R^(2a))(CH₂)₀₋₄OR^(3a),    -   (3) —(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a),R^(5a)),    -   (4) —(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a))COR^(3a),    -   (5) —(CH₂)₀₋₄C(R^(1a),R^(2a))NHCON(R^(4a),R^(5a)),    -   (6) —(CH₂)₀₋₄C(R^(1a),R^(2a))NHC(NH)N(R^(4a),R^(5a)),    -   (7) —CH(R^(1a),R^(2a)),    -   (8) —C≡CH,    -   (9) —(CH₂)₀₋₄C(R^(1a),R^(2a))CN,    -   (10) —(CH₂)₀₋₄C(R^(1a),R^(2a))CO₂R^(3a),    -   (11) —(CH₂)₀₋₄C(R^(1a),R^(2a))CON(R^(4a),R^(5a)),    -   (12) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (13) substituted or unsubstituted aryl,    -   (14) substituted or unsubstituted heterocyclyl, and    -   (15) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1a), R^(2a), R^(3a), R^(4a), and R^(5a) is            independently selected from the group consisting of:            -   (a) H,            -   (b) a halogen atom,            -   (c) substituted or unsubstituted C₁-C₆-alkyl,            -   (d) substituted or unsubstituted aryl,            -   (e) substituted or unsubstituted heterocyclyl, and            -   (f) substituted or unsubstituted heteroaryl,    -   or R^(4a) and R^(5a) together with the N atom to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 5 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S;

Q is absent or selected from the group consisting of:

-   -   (1) —C(═O)N(R₁,R₂),    -   (2) —NHC(═O)N(R₁,R₂),    -   (3) —N(OH)C(═O)N(R₁,R₂),    -   (4) —CH(OH)C(═O)N(R₁,R₂),    -   (5) —CH[N(R^(2q), R^(3q))]C(═O)N(R₁,R₂),    -   (6) —CHR^(1q)C(═O)N(R₁, R₂),    -   (7) —CO₂H,    -   (8) —C(═O)NHSO₂R^(4q),    -   (9) —SO₂NH₂,    -   (10) —N(OH)C(═O)R^(1q),    -   (11) —N(OH)SO₂R^(4q),    -   (12) —NHSO₂R^(4q),    -   (13) —SH,    -   (14) —CH(SH)(CH₂)₀₋₁C(═O)N(R₁,R₂),    -   (15) —CH(SH)(CH₂)₀₋₁CO₂H,    -   (16) —CH(OH)(CH₂)₀₋₁CO₂H,    -   (17) —CH(SH)CH₂CO₂R^(1q),    -   (18) —CH(OH)(CH₂)SO₂NH₂,    -   (19) —CH(CH₂SH)NHCOR^(1q),    -   (20) —CH(CH₂SH)NHSO₂R^(4q),    -   (21) —CH(CH₂SR^(5q))CO₂H,    -   (22) —CH(CH₂SH)NHSO₂NH₂,    -   (23) —CH(CH₂OH)CO₂H,    -   (24) —CH(CH₂OH)NHSO₂NH₂,    -   (25) —C(═O)CH₂CO₂H,    -   (26) —C(═O)(CH₂)₀₋₁CONH₂,    -   (27) —OSO₂NHR^(5q),    -   (28) —SO₂NHNH₂,    -   (29) —P(═O)(OH)₂,

-   -   and    -   (33) —N(OH)C(═O)CR₁R₂,    -   wherein:        -   R₁ is selected from the group consisting of:            -   (1) —H,            -   (2) —OH,            -   (3) —OC₁-C₆-alkyl,            -   (4) —N(R^(2q), R^(3q)), and            -   (5) substituted or unsubstituted C₁-C₆-alkyl;        -   R₂ is selected from the group consisting of:            -   (1) H,            -   (2) substituted or unsubstituted C₁-C₆-alkyl;            -   (3) substituted or unsubstituted C₂-C₆-alkenyl,            -   (4) substituted or unsubstituted C₂-C₆-alkenyl,            -   (5) substituted or unsubstituted aryl,            -   (6) substituted or unsubstituted heterocyclyl, and            -   (7) substituted or unsubstituted heteroaryl,        -   or R₁ and R₂, together with the N atom to which they are            attached can form a substituted or unsubstituted            heterocyclic ring, having from 3 to 10 ring atoms, wherein            1-4 ring atoms of the heterocyclic ring are selected from N,            O and S; and        -   each R^(1q), R^(2q), R^(3q), R^(4q), and R^(5q) is            independently selected from the group consisting of H and            C₁-C₆ alkyl.

In a second embodiment, the present invention provides compounds offormula (I) wherein:

E is selected from the group consisting of:

-   -   (1) H,    -   (2) substituted or unsubstituted C₁-C₆-alkyl,    -   (3) substituted or unsubstituted C₂-C₆-alkenyl,    -   (4) substituted or unsubstituted C₂-C₆-alkynyl,    -   (5) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (6) substituted or unsubstituted aryl,    -   (7) substituted or unsubstituted heterocyclyl, and    -   (8) substituted or unsubstituted heteroaryl;

L is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₁-C₆-alkyl,    -   (2) —(NR^(3L))₀₋₁—(CH₂)₀₋₄—NR^(3L)—(CH₂)₀₋₄—,    -   (3) —(NR^(3L))₀₋₁—C(R^(1L),R^(2L))—NR^(3L)—C(R^(1L),R^(2L))—,    -   (4) —C(R^(1L),R^(2L))—O—C(R^(1L),R^(2L))—,    -   (5) —(CH₂)₀₋₄—NR^(3L)—C(R^(1L),R^(2L))—CONH—(CH₂)₀₋₄—,    -   (6) —CO—C(R^(1L),R^(2L))—NHCO—,    -   (7) —CONR^(3L)—,    -   (8) —NR^(3L)CO—,    -   (9) —NR^(3L)—,    -   (10) —SO₂NR^(3L)—,    -   (11) —NR^(3L)—C(═O)—NR^(3L)—,    -   (12) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (13) substituted or unsubstituted aryl,    -   (14) substituted or unsubstituted heterocyclyl, and    -   (15) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1L), R^(2L), and R^(3L) is independently selected            from the group consisting of:            -   (a) H,            -   (b) substituted or unsubstituted C₁-C₆-alkyl,            -   (c) C₁-C₆-alkyl substituted with aryl,            -   (d) C₁-C₆-alkyl substituted with heterocyclyl, and            -   (e) C₁-C₆-alkyl substituted with heteroaryl,    -   or R^(1L) and R^(3L), together with the atoms to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 3 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S;

D is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

G is selected from the group consisting of:

-   -   (1) —(CH₂)₀₋₄—O—(CH₂)₀₋₄—,    -   (2) —(CH₂)₀₋₄—S—(CH₂)₀₋₄—,    -   (3) —(CH₂)₀₋₄—NR^(1G)—(CH₂)₀₋₄—,    -   (4) —C(═O)—,    -   (5) —NR^(1G)C(═O)—,    -   (6) —C(═O)NR^(1G)—,    -   (7) —(CH₂)₀₋₄NHCH₂C(═O)NR^(1G)—,    -   (8) —C≡C—,    -   (9) —C≡C—C≡C—,    -   (10) —CR^(2G)═CR^(2G)—,    -   (11) —S(═O)—,    -   (12) —SO₂—,    -   (13) —C(R^(3G))₂—S(═O)—,    -   (14) —S(═O)—C(R^(3G))₂—,    -   (15) —C(R^(3G))₂—SO₂—,    -   (16) —SO₂—C(R^(3G))₂—,    -   (17) —CR^(3G)═CR^(3G)—CR^(3G)═CR^(3G)—,    -   (18) —C(R^(3G))₂—,    -   (19) —CR^(3G)═CR^(3G)—C≡C—,    -   (20) —C≡C—CR^(3G)═CR^(3G)—,    -   (21) —C(═O)—C≡C—,    -   (22) —C≡C—C(═O)—,    -   (23) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (24) substituted or unsubstituted aryl,    -   (25) substituted or unsubstituted heterocyclyl, and    -   (26) substituted or unsubstituted heteroaryl,    -   wherein:        -   R^(1G) is substituted or unsubstituted C₁-C₆-alkyl;        -   each R^(2G) and R^(3G) is independently selected from the            group consisting of H, a halogen atom, and substituted or            unsubstituted C₁-C₆-alkyl;

Y is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

X is selected from the group consisting of:

-   -   (1) —(C═O)NR₄—,    -   (2) —C₁-C₆-alkyl-(C═O)NR₄—,    -   (3) —C₂-C₆-alkenyl-(C═O)NR₄—,    -   (4) —C₂-C₆-alkynyl-(C═O)NR₄—,    -   (5) —CH₂NR₄—,    -   (6) —SO₂NR₄—,    -   (7) —S(═O)NR₄—,    -   (8) —NR₄C(═O)—, and    -   (9) —NR₄—,    -   or X and A, together with the atoms to which they are attached        can form a heterocyclic ring, having from 5 to 8 ring atoms,        wherein 1-2 ring atoms of the heterocyclic ring are selected        from N, O and S,    -   or when Y is a bicyclic substituted or unsubstituted        heterocyclyl or heteroaryl, then X is absent;

R₃ is H or substituted or unsubstituted C₁-C₆-alkyl, or R₃ and A,together with the atom to which they are attached can form a substitutedor unsubstituted 3-10 membered cycloalkyl or a heterocyclic ring, havingfrom 3 to 10 ring atoms, wherein 1-4 ring atoms of the heterocyclic ringare selected from N, O and S;

R₄ is (1) H or substituted or unsubstituted C₁-C₆-alkyl, or (2) R₄ andA, together with the atoms to which they are attached can form asubstituted or unsubstituted heterocyclic ring, having from 3 to 8 ringatoms, wherein 1-2 ring atoms of the heterocyclic ring are selected fromN, O and S, or (3) R₄ and Y, together with the atoms to which they areattached, form a bicyclic substituted or unsubstituted heterocyclyl orheteroaryl;

n is an integer from 0-6;

A is selected from the group consisting of:

-   -   (1) —C(R^(1a),R^(2a))OR^(3a),    -   (2) —C(R^(1a),R^(2a))N(R^(4a),R^(5a)),    -   (3) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (4) substituted or unsubstituted aryl,    -   (5) substituted or unsubstituted heterocyclyl, and    -   (6) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1a) and R^(2a) is independently selected from the            group consisting of substituted or unsubstituted            C₁-C₆-alkyl;        -   each R^(1a), R^(4a), and R^(5a) is independently selected            from the group consisting of:            -   (a) H,            -   (b) a halogen atom,            -   (c) substituted or unsubstituted C₁-C₆-alkyl,            -   (d) substituted or unsubstituted aryl,            -   (e) substituted or unsubstituted heterocyclyl, and            -   (f) substituted or unsubstituted heteroaryl,    -   or R^(4a) and R^(Sa) together with the N atom to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 5 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S; and        -   when A is —C(R^(1a),R^(2a))OR^(3a), the compound is not            2-{[(4′-ethyl-1,1′-biphenyl-4-yl)carbonyl]amino}-3-hydroxy-3-methylbutanoic            acid, 4′-ethyl-N-{2-hydroxy-1-[(hydroxyamino)            carbonyl]-2-methylpropyl}-1,1′-biphenyl-4-carboxamide or            N-{2-hydroxy-1-[(hydroxyamino)carbonyl]-2-methylpropyl}-4-(phenylethynyl)benzamide;

Q is absent or selected from the group consisting of:

-   -   (1) —C(═O)N(R₁,R₂),    -   (2) —NHC(═O)N(R₁,R₂),    -   (3) —N(OH)C(═O)N(R₁,R₂),    -   (4) —CH(OH)C(═O)N(R₁,R₂),    -   (5) —CH[N(R^(2q), R^(3q))]C(═O)N(R₁,R₂),    -   (6) —CHR^(1q)C(═O)N(R₁, R₂),    -   (7) —CO₂H,    -   (8) —C(═O)NHSO₂R^(4q),    -   (9) —SO₂NH₂,    -   (10) —N(OH)C(═O)R^(1q),    -   (11) —N(OH)SO₂R^(4q),    -   (12) —NHSO₂R^(4q),    -   (13) —SH,    -   (14) —CH(SH)(CH₂)₀₋₁C(═O)N(R₁,R₂),    -   (15) —CH(SH)(CH₂)₀₋₁CO₂H,    -   (16) —CH(OH)(CH₂)₀₋₁CO₂H,    -   (17) —CH(SH)CH₂CO₂R^(1q),    -   (18) —CH(OH)(CH₂)SO₂NH₂,    -   (19) —CH(CH₂SH)NHCOR^(1q),    -   (20) —CH(CH₂SH)NHSO₂R^(4q),    -   (21) —CH(CH₂SR^(5q))CO₂H,    -   (22) —CH(CH₂SH)NHSO₂NH₂,    -   (23) —CH(CH₂OH)CO₂H,    -   (24) —CH(CH₂OH)NHSO₂NH₂,    -   (25) —C(═O)CH₂CO₂H,    -   (26) —C(═O)(CH₂)₀₋₁CONH₂,    -   (27) —OSO₂NHR^(5q),    -   (28) —SO₂NHNH₂,    -   (29) —P(═O)(OH)₂,

-   -   and    -   (33) —N(OH)C(═O)CR₁R₂,    -   wherein:        -   R₁ is selected from the group consisting of:            -   (1) —H,            -   (2) —OH,            -   (3) —OC₁-C₆-alkyl,            -   (4) —N(R^(2q), R^(3q)), and            -   (5) substituted or unsubstituted C₁-C₆-alkyl;        -   R₂ is selected from the group consisting of:            -   (1) H,            -   (2) substituted or unsubstituted C₁-C₆-alkyl,            -   (3) substituted or unsubstituted C₂-C₆-alkenyl,            -   (4) substituted or unsubstituted C₂-C₆-alkenyl,            -   (5) substituted or unsubstituted aryl,            -   (6) substituted or unsubstituted heterocyclyl, and            -   (7) substituted or unsubstituted heteroaryl,        -   or R₁ and R₂, together with the N atom to which they are            attached can form a substituted or unsubstituted            heterocyclic ring, having from 3 to 10 ring atoms, wherein            1-4 ring atoms of the heterocyclic ring are selected from N,            O and S; and        -   each R^(1q), R^(2q), R^(3q), R^(4q), and R^(5q) is            independently selected from the group consisting of H and            C₁-C₆ alkyl.

In another aspect, the present invention provides a pharmaceuticalcomposition comprising a compound of formula (I) and a pharmaceuticallyacceptable carrier or diluent.

In another aspect, the present invention provides a pharmaceuticalcomposition or formulation comprising an effective amount of a compoundof formula (I) and a pharmaceutically acceptable carrier or diluent.

In another aspect, the present invention provides a method of inhibitinga deacetylase enzyme in gram-negative bacteria, thereby affectingbacterial growth, comprising administering to a patient in need of suchinhibition a compound of formula (I).

In another aspect, the present invention provides a method of inhibitingLpxC, thereby modulating the virulence of a bacterial infection,comprising administering to a patient in need of such inhibition acompound of formula (I).

In another aspect, the present invention provides a method for treatinga subject with a gram-negative bacterial infection comprisingadministering to the subject in need thereof an antibacteriallyeffective amount of a compound of formula (I) with a pharmaceuticallyacceptable carrier. In a more specific embodiment of the method oftreatment, the subject is a mammal and in certain embodiments, a human.

In another aspect, the present invention provides a method ofadministering an inhibitory amount of a compound of formula (I) to asubject infected with a fermentative or non-fermentative gram-negativebacteria. In a more specific embodiment of the method of administeringan inhibitory amount of a compound of formula (I) to a subject infectedwith a fermentative or non-fermentative gram-negative bacteria, thegram-negative bacteria are selected from the group consisting ofPseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderiacepacia, Alcaligenes xylosoxidans, Acinetobacter, Enterobacteriaceae,Haemophilus, Franciscellaceae (Franciscella tularensis) and Neisseriaspecies.

In another aspect, the present invention provides a method ofadministering an inhibitory amount of a compound of formula (I) to asubject infected with gram-negative bacteria, such as a member of theEnterobacteriaceae which is selected from the group consisting oforganisms such as Serratia, Proteus, Klebsiella, Enterobacter,Citrobacter, Salmonella, Providencia, Yersinia (Yersinia pestis),Morganella, Cedecea, and Edwardsiella species and Escherichia coli.

In another aspect, the present invention provides a method ofco-administering a compound of formula (I) with other therapeutic agentsthat are selected for their particular usefulness against the conditionthat is being treated. For example, a compound of formula (I) is usefulin combination with other anti-bacterial agents. The compound of formula(I) augments the sensitivity of gram-negative bacteria to existingclasses of antibacterials. Combinations of the presently disclosedcompounds with other anti-bacterial agents are within the scope of theinvention. Such anti-bacterial agents include, but are not limited to,erythromycin, rifampicin, Nalidixic acid, carbenicillin, bacitracin,cycloserine, fosfomycin, and vancomycin.

These and other aspects of the invention will be evident upon referenceto the following detailed description.

DETAILED DESCRIPTION OF THE INVENTION

Unless the context requires otherwise, throughout the specification andclaims which follow, the word “comprise” and variations thereof, suchas, “comprises” and “comprising” are to be construed in an open,inclusive sense, that is as “including, but not limited to”.

Reference throughout this specification to “one embodiment” or “anembodiment” means that a particular feature, structure or characteristicdescribed in connection with the embodiment is included in at least oneembodiment of the present invention. Thus, the appearances of thephrases “in one embodiment” or “in an embodiment” in various placesthroughout this specification are not necessarily all referring to thesame embodiment. Furthermore, the particular features, structures, orcharacteristics may be combined in any suitable manner in one or moreembodiments.

The present invention provides novel compounds, methods for inhibitingLpxC in gram-negative bacteria, and novel methods for treating bacterialinfections. The compounds provided herein can be formulated intopharmaceutical formulations and medicaments that are useful in themethods of the invention. The invention also provides for the use of thecompounds in preparing medicaments and pharmaceutical formulations, foruse of the compounds in inhibiting LpxC, and for use of the compounds intreating bacterial infections in a subject.

The following abbreviations and definitions are used throughout thisapplication:

“LpxC” is an abbreviation that stands forUDP-3-O—(R-3-hydroxydecanoyl)-N-acetylglucosamine deacetylase.

Generally, reference to a certain element such as hydrogen or H is meantto include all isotopes of that element. For example, if a substituentgroup is defined to include hydrogen or H, it also includes deuteriumand tritium.

“Alkyl” refers to a straight or branched, saturated hydrocarbon chainradical consisting solely of carbon and hydrogen atoms, and having fromone to twelve carbon atoms, preferably one to eight carbon atoms(C₁-C₈-alkyl) or one to six carbon atoms (C₁-C₆-alkyl). Thus the phraseincludes straight chain alkyl groups such as methyl, ethyl, propyl,butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl andthe like. The phrase also includes branched chain isomers of straightchain alkyl groups, including but not limited to, the following, thatare provided by way of example: —CH(CH₃)₂, —CH(CH₃)(CH₂CH₃),—CH(CH₂CH₃)₂, —C(CH₃)₃, —C(CH₂CH₃)₃, —CH₂CH(CH₃)₂, —CH₂CH(CH₃)(CH₂CH₃),—CH₂CH(CH₂CH₃)₂, —CH₂C(CH₃)₃, —CH₂C(CH₂CH₃)₃, —CH(CH₃)CH(CH₃)(CH₂CH₃),—CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH(CH₃)(CH₂CH₃), —CH₂CH₂CH(CH₂CH₃)₂,—CH₂CH₂C(CH₃)₃, —CH₂CH₂C(CH₂CH₃)₃, —CH(CH₃)CH₂CH(CH₃)₂,—CH(CH₃)CH(CH₃)CH(CH₃)₂, —CH(CH₂CH₃)CH(CH₃)CH(CH₃)(CH₂CH₃) and the like.

“Alkenyl” refers to a straight or branched, unsaturated hydrocarbonchain radical containing at least one double bond, consisting solely ofcarbon and hydrogen atoms and having from two to twelve carbon atoms,preferably two to eight carbon atoms (C₂-C₈-alkenyl) or two to sixcarbon atoms (C₂-C₆-alkenyl). Representative alkenyl radicals include,but are not limited to, vinyl, —CH═CH(CH₃), —CH═C(CH₃)₂, —C(CH₃)═CH₂,—C(CH₃)═CH(CH₃), —C(CH₂CH₃)═CH₂, cyclohexenyl, cyclopentenyl,cyclohexadienyl, butadienyl, pentadienyl, hexadienyl and the like.

“Alkynyl” refers to a straight or branched, unsaturated hydrocarbonchain radical containing at least one triple bond, and, optionally, atleast one double bond, consisting solely of carbon and hydrogen atomsand having from two to twelve carbon atoms, preferably two to eightcarbon atoms (C₂-C₈-alkynyl) or two to six carbon atoms (C₂-C₆-alkynyl).Representative alkynyl radicals include, but are not limited to —C≡CH,—C≡C(CH₃), —C≡C(CH₂CH₃), —CH₂C≡CH, —CH₂C≡C(CH₃), —CH₂C≡C(CH₂CH₃) and thelike.

“Alkoxy” refers to a radical of the formula —OR_(a) where R_(a) is analkyl, alkenyl or alkynyl radical as defined above containing one totwelve carbon atoms. Unless stated otherwise specifically in thespecification, an alkoxy group may be optionally substituted.

“Alkylamino” refers to a radical of the formula —NHR_(a) or —NR_(a)R_(a)where each R_(a) is, independently, an alkyl, alkenyl or alkynyl radicalas defined above containing one to twelve carbon atoms. Unless statedotherwise specifically in the specification, an alkylamino group may beoptionally substituted.

“Thioalkyl” refers to a radical of the formula —SR_(a) where R_(a) is analkyl, alkenyl or alkynyl radical as defined above containing one totwelve carbon atoms. Unless stated otherwise specifically in thespecification, a thioalkyl group may be optionally substituted.

“Aryl” refers to aromatic monocyclic or multicyclic hydrocarbon ringsystems consisting only of hydrogen and carbon atoms and containing from6 to 19 carbon atoms, where the ring system may be partially or fullysaturated, e.g., phenyl, biphenyl, anthracenyl, naphthyl, and the like.

“Cycloalkyl” refers to a non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,which may include fused or bridged ring systems, having from three tofifteen carbon atoms, preferably having from three to ten carbon atoms(C₃-C₁₀-cycloalkyl), and which is saturated or unsaturated. Monocyclicradicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptly, cyclooctyl, and the like. Polycyclic radicalsinclude, for example, adamantine, norbornane, decalinyl,7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.

“Heteroaryl” or “heteroaryl ring” refers to a 5- to 18-membered aromaticring radical which consists of three to seventeen carbon atoms and fromone to ten heteroatoms selected from the group consisting of nitrogen,oxygen and sulfur. For purposes of this invention, the heteroarylradical may be a monocyclic, bicyclic or polycyclic ring system, whichmay include fused or bridged ring systems; the nitrogen, carbon orsulfur atoms in the heteroaryl radical may be optionally oxidized; andthe nitrogen atom may be optionally quaternized. Heteroaryl radicalsinclude, for example, azepinyl, acridinyl, benzimidazolyl,benzthiazolyl, be nzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl,benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl,1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl,benzo-1,3-dioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl,benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl),benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl,cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl,isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl,indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyl,naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl,phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl,pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl,quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thiophenyl(i.e., thienyl), and the like.

“Heterocyclic ring” includes both “heterocyclyl” and “heteroaryl”groups.

“Heterocyclyl” refers to a 3- to 18-membered non-aromatic ring radicalwhich consists of two to seventeen carbon atoms and from one to tenheteroatoms selected from the group consisting of nitrogen, oxygen andsulfur. Unless stated otherwise specifically in the specification, theheterocyclyl radical may be a monocyclic, bicyclic or polycyclic ringsystem, which may include fused or bridged ring systems; and thenitrogen, carbon or sulfur atoms in the heterocyclyl radical may beoptionally oxidized; the nitrogen atom may be optionally quaternized;and the heterocyclyl radical may be partially or fully saturated.Heterocyclyl radicals include, for example, dioxolanyl,thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, oxiranyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl,1,1-dioxo-thiomorpholinyl, and the like.

Representative heteroaryl and heterocyclyl radicals include, but are notlimited to: unsaturated 3 to 8 membered rings containing 1 to 4 nitrogenatoms such as, but not limited to pyrrolyl, pyrrolinyl, imidazolyl,pyrazolyl, pyridyl, dihydropyridyl, pyrimidyl, pyrazinyl, pyridazinyl,triazolyl (e.g. 4H-1,2,4-triazolyl, 1H-1,2,3-triazolyl,2H-1,2,3-triazolyl etc.), tetrazolyl, (e.g. 1H-tetrazolyl, 2Htetrazolyl, etc.); saturated 3 to 8 membered rings containing 1 to 4nitrogen atoms such as, but not limited to, pyrrolidinyl,imidazolidinyl, piperidinyl, piperazinyl; condensed unsaturatedheterocyclic groups containing 1 to 4 nitrogen atoms such as, but notlimited to, indolyl, isoindolyl, indolinyl, indolizinyl, benzimidazolyl,quinolyl, isoquinolyl, indazolyl, benzotriazolyl; unsaturated 3 to 8membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atomssuch as, but not limited to, oxazolyl, isoxazolyl, oxadiazolyl (e.g.1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.);saturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to3 nitrogen atoms such as, but not limited to, morpholinyl; unsaturatedcondensed heterocyclic groups containing 1 to 2 oxygen atoms and 1 to 3nitrogen atoms, for example, benzoxazolyl, benzoxadiazolyl, benzoxazinyl(e.g. 2H-1,4-benzoxazinyl etc.); unsaturated 3 to 8 membered ringscontaining 1 to 3 sulfur atoms and 1 to 3 nitrogen atoms such as, butnot limited to, thiazolyl, isothiazolyl, thiadiazolyl (e.g.1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl,1,2,5-thiadiazolyl, etc.); saturated 3 to 8 membered rings containing 1to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to,thiazolodinyl; saturated and unsaturated 3 to 8 membered ringscontaining 1 to 2 sulfur atoms such as, but not limited to, thienyl,dihydrodithiinyl, dihydrodithionyl, tetrahydrothiophene,tetrahydrothiopyran; unsaturated condensed heterocyclic rings containing1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limitedto, benzothiazolyl, benzothiadiazolyl, benzothiazinyl (e.g.2H-1,4-benzothiazinyl, etc.), dihydrobenzothiazinyl (e.g.2H-3,4-dihydrobenzothiazinyl, etc.), unsaturated 3 to 8 membered ringscontaining oxygen atoms such as, but not limited to furyl; unsaturatedcondensed heterocyclic rings containing 1 to 2 oxygen atoms such asbenzodioxolyl (e.g. 1,3-benzodioxoyl, etc.); unsaturated 3 to 8 memberedrings containing an oxygen atom and 1 to 2 sulfur atoms such as, but notlimited to, dihydrooxathiinyl; saturated 3 to 8 membered ringscontaining 1 to 2 oxygen atoms and 1 to 2 sulfur atoms such as1,4-oxathiane; unsaturated condensed rings containing 1 to 2 sulfuratoms such as benzothienyl, benzodithiinyl; and unsaturated condensedheterocyclic rings containing an oxygen atom and 1 to 2 oxygen atomssuch as benzoxathiinyl. Heterocyclyl group also include those describedabove in which one or more S atoms in the ring is double-bonded to oneor two oxygen atoms (sulfoxides and sulfones). For example, heterocyclylgroups include tetrahydrothiophene, tetrahydrothiophene oxide, andtetrahydrothiophene 1,1-dioxide. Preferred heterocyclyl groups contain 5or 6 ring members. More preferred heterocyclyl groups includemorpholine, piperazine, piperidine, pyrrolidine, imidazole, pyrazole,1,2,3-triazole, 1,2,4-triazole, tetrazole, thiomorpholine,thiomorpholine in which the S atom of the thiomorpholine is bonded toone or more 0 atoms, pyrrole, homopiperazine, oxazolidin-2-one,pyrrolidin-2-one, oxazole, quinuclidine, thiazole, isoxazole, furan, andtetrahydrofuran.

The term “substituted” used herein means any of the above groups (i.e.,alkyl, alkenyl, alkynyl, alkylamino, alkoxy, alkylthio, aryl,cycloalkyl, heteroaryl and/or heterocyclyl) wherein at least onehydrogen atom is replaced by a bond to a non-hydrogen atoms such as, butnot limited to: a halogen atom such as F, Cl, Br, and I; an oxygen atomin groups such as hydroxyl groups, alkoxy groups, and ester groups; asulfur atom in groups such as thiol groups, alkyl and aryl sulfidegroups, sulfone groups, sulfonyl groups, and sulfoxide groups; anitrogen atom in groups such as amines, amides, alkylamines,dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides,imides, and enamines; a silicon atom in groups such as trialkylsilylgroups, dialkylarylsilyl groups, alkyldiarylsilyl groups, andtriarylsilyl groups; and other heteroatoms in various other groups.“Substituted” also means any of the above groups in which one or morebonds are replaced by a higher-order bond (e.g., a double- ortriple-bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl,and ester groups; and nitrogen in groups such as imines, oximes,hydrazones, and nitriles. “Substituted” further means any of the abovegroups in which one or more bonds are replaced by a bond to an alkyl,alkenyl, alkynyl, amino, aryl, cyano, cycloalkyl, halogen, heteroaryl,heterocyclyl, hydroxyl, imino, nitro, oxo or thioxo group. For example,“substituted” includes any of the above groups in which one or morehydrogen atoms are replaced with —NR_(g)R_(h), —NR_(g)C(═O)R_(h),—NR_(g)C(═O)NR_(g)R_(h), —NR_(g)C(═O)OR_(h), —NR_(g)SO₂R_(h),—OC(═O)NR_(g)R_(h), —OR_(g), —SR_(g), —SOR_(g), —SO₂R_(g), —OSO₂R_(g),—SO₂OR_(g), ═NSO₂R_(g), and —SO₂NR_(g)R_(h). “Substituted” also meansany of the above groups in which one or more hydrogen atoms are replacedwith —C(═O)R_(g), —C(═O)OR_(g), —C(═O)NR_(g)R_(h), —CH₂SO₂R_(g),—CH₂SO₂NR_(g)R_(h). In the foregoing, R_(g) and R_(h) are the same ordifferent and independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy,alkylamino, alkylthio, aryl, cycloalkyl, heterocyclyl and/or heteroaryl.In addition, the foregoing substituents may also be optionallysubstituted with one or more of the above substituents.

For example, representative substituted alkyl groups includetrifluoromethyl. Representative heteroaryl and heterocyclyl groupsinclude 2-methylbenzimidazolyl, 5-methylbenzimidazolyl,5-chlorobenzthiazolyl, 1-methylpiperazinyl, and 2-chloropyridyl.Representative substituted aryl groups include tolyl and hydroxyphenyl.Other representative substitutents include straight and branched chainalkyl groups, —CH₃, —C₂H₅, —CH₂OH, —OH, —OCH₃, —OC₂H₅, —OCF₃, —CN, —NO₂,—CO₂H, —CO₂CH₃, —CONH₂, —NH₂, —F, —Cl, —Br, —CF₃, —N(CH₃)₂, —NHSO₂CH₃,and —NHCOCH₃.

“Amino” refers to the —NH₂ radical.

“Cyano” refers to the —CN radical.

“Halogen” refers to bromo, chloro, fluoro or iodo.

“Hydroxyl” refers to the —OH radical.

“Imino” refers to the ═NH substituent.

“Nitro” refers to the —NO₂ radical.

“Oxo” refers to the ═O substituent.

“Thioxo” refers to the ═S substituent.

“Fused” refers to any ring structure described herein which is fused toan existing ring structure in the compounds of the present invention.When the fused ring is a heterocyclyl ring or a heteroaryl ring, anycarbon atom on the existing ring structure which becomes part of thefused heterocyclyl ring or the fused heteroaryl ring may be replacedwith a nitrogen atom.

The term “protected” with respect to hydroxyl groups, amine groups, andsulfhydryl groups refers to forms of these functionalities that areprotected from undesirable reaction with a protecting group known tothose skilled in the art such as those set forth in Protective Groups inOrganic Synthesis, Greene, T. W.; Wuts, P. G. M., John Wiley & Sons, NewYork, N.Y., (3rd Edition, 1999) that can be added or removed using theprocedures set forth therein. Examples of protected hydroxyl groupsinclude, but are not limited to, silyl ethers such as those obtained byreaction of a hydroxyl group with a reagent such as, but not limited to,t-butyldimethyl-chlorosilane, trimethylchlorosilane,triisopropylchlorosilane, triethylchlorosilane; substituted methyl andethyl ethers such as, but not limited to methoxymethyl ether,methythiomethyl ether, benzyloxymethyl ether, t-butoxymethyl ether,2-methoxyethoxymethyl ether, tetrahydropyranyl ethers, 1-ethoxyethylether, allyl ether, benzyl ether; esters such as, but not limited to,benzoylformate, formate, acetate, trichloroacetate, and trifluoracetate.Examples of protected amine groups include, but are not limited to,amides such as, formamide, acetamide, trifluoroacetamide, and benzamide;imides, such as phthalimide, and dithiosuccinimide; and others. Examplesof protected sulfhydryl groups include, but are not limited to,thioethers such as S-benzyl thioether, and S-4-picolyl thioether;substituted S-methyl derivatives such as hemithio, dithio and aminothioacetals; and others.

The term “pharmaceutically acceptable salt” includes a salt with aninorganic base, organic base, inorganic acid, organic acid, or basic oracidic amino acid. As salts of inorganic bases, the invention includes,for example, alkali metals such as sodium or potassium; alkaline earthmetals such as calcium and magnesium or aluminum; and ammonia. As saltsof organic bases, the invention includes, for example, trimethylamine,triethylamine, pyridine, picoline, ethanolamine, diethanolamine, andtriethanolamine. As salts of inorganic acids, the instant inventionincludes, for example, hydrochloric acid, hydroboric acid, nitric acid,sulfuric acid, and phosphoric acid. As salts of organic acids, theinstant invention includes, for example, formic acid, acetic acid,trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, maleicacid, citric acid, succinic acid, malic acid, methanesulfonic acid,benzenesulfonic acid, and p-toluenesulfonic acid. As salts of basicamino acids, the instant invention includes, for example, arginine,lysine and ornithine. Acidic amino acids include, for example, asparticacid and glutamic acid.

The term “pharmaceutically acceptable ester” refers to esters thathydrolyze in vivo and include those that break down readily in the humanbody to leave the parent compound or a salt thereof. Suitable estergroups include, for example, those derived from pharmaceuticallyacceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic,cycloalkanoic and alkanedioic acids, in which each alkyl or alkenylmoiety advantageously has not more than 6 carbon atoms. Representativeexamples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrug” as used herein refers tothose prodrugs of the compounds of the present invention that are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals without undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of theinvention. The term “prodrug” refers to compounds that are rapidlytransformed in vivo to yield the parent compound of the above formula,for example by hydrolysis in blood. A thorough discussion is provided inT. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14of the A.C.S. Symposium Series, and in Edward B. Roche, ed.,Bioreversible Carriers in Drug Design, American PharmaceuticalAssociation and Pergamon Press, 1987, both of which are incorporatedherein by reference.

The term “antibacterial agent” refers to agents synthesized or modifiedin the laboratory that have either bactericidal or bacteriostaticactivity. An “active” agent in this context will inhibit the growth ofP. aeruginosa and other gram-negative bacteria. The term “inhibiting thegrowth” indicates that the rate of increase in the numbers of apopulation of a particular bacterium is reduced. Thus, the term includessituations in which the bacterial population increases but at a reducedrate, as well as situations where the growth of the population isstopped, as well as situations where the numbers of the bacteria in thepopulation are reduced or the population even eliminated. If an enzymeactivity assay is used to screen for inhibitors, one can makemodifications in uptake/efflux, solubility, half-life, etc. to compoundsin order to correlate enzyme inhibition with growth inhibition. Theactivity of antibacterial agents is not necessarily limited to bacteriabut may also encompass activity against parasites, virus, and fungi.

The subject invention also includes isotopically-labeled LpxCinhibitors, that are structurally identical to those disclosed herein,but for the fact that one or more atoms are replaced by an atom havingan atomic mass or mass number different from the atomic mass or massnumber usually found in nature. Examples of isotopes that can beincorporated into compounds of the invention include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine andchlorine, such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸Fand ³⁶Cl, respectively. Compounds of the present invention, prodrugsthereof, and pharmaceutically acceptable salts of said compounds and ofsaid prodrugs that contain the aforementioned isotopes and/or otherisotopes of other atoms are within the scope of this invention. Certainisotopically labeled compounds of the present invention, for examplethose into which radioactive isotopes such as ³H and ¹⁴C areincorporated, are useful in drug and/or substrate tissue distributionassays. Tritiated, i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes areparticularly preferred for their ease of preparation and detectability.Further, substitution with heavier isotopes such as deuterium, i.e., ²H,may afford certain therapeutic advantages resulting from greatermetabolic stability, for example increased in vivo half-life or reduceddosage requirements and, hence, may be preferred in some circumstances.Isotopically labeled compounds of this invention and prodrugs thereofcan generally be prepared by carrying out known or referenced proceduresand by substituting a readily available isotopically labeled reagent fora non-isotopically labeled reagent.

As noted above, in one aspect, the present invention provides compoundshaving the following formula (I):

including stereoisomers, pharmaceutically acceptable salts, esters, andprodrugs thereof.

In a first embodiment, the present invention provides compounds offormula (I) wherein:

E is selected from the group consisting of:

-   -   (1) H,    -   (2) substituted or unsubstituted C₁-C₆-alkyl,    -   (3) substituted or unsubstituted C₂-C₆-alkenyl,    -   (4) substituted or unsubstituted C₂-C₆-alkynyl,    -   (5) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (6) substituted or unsubstituted aryl,    -   (7) substituted or unsubstituted heterocyclyl, and    -   (8) substituted or unsubstituted heteroaryl;

L is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₁-C₆-alkyl,    -   (2) —(NR^(3L))₀₋₁—(CH₂)₀₋₄—NR^(3L)—(CH₂)₀₋₄—,    -   (3) —(NR^(3L))₀₋₁—C(R^(1L),R^(2L))—NR^(3L) C(R^(1L),R^(2L))—,    -   (4) —C(R^(1L),R^(2L))—O—C(R^(1L),R^(2L))—,    -   (5) —(CH₂)₀₋₄—NR^(3L)—C(R^(1L),R^(2L))—CONH—(CH₂)₀₋₄—,    -   (6) —CO—C(R^(1L),R^(2L))—NHCO—,    -   (7) —CONR^(3L)—,    -   (8) —NR^(3L)CO—,    -   (9) —NR^(3L)—,    -   (10) —SO₂NR^(3L)—,    -   (11) —NR^(3L)—C(═O)—NR^(3L)—,    -   (12) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (13) substituted or unsubstituted aryl,    -   (14) substituted or unsubstituted heterocyclyl, and    -   (15) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1L), R^(2L), and R^(3L) is independently selected            from the group consisting of:            -   (a) H,            -   (b) substituted or unsubstituted C₁-C₆-alkyl,            -   (c) C₁-C₆-alkyl substituted with aryl,            -   (d) C₁-C₆-alkyl substituted with heterocyclyl, and            -   (e) C₁-C₆-alkyl substituted with heteroaryl,    -   or R^(1L) and R^(3L), together with the atoms to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 3 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S;

D is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

G is selected from the group consisting of:

-   -   (1) —NR^(1G)C(═O)—,    -   (2) —C(═O)NR^(1G)—,    -   (3) —(CH₂)₀₋₄NHCH₂C(═O)NR^(1G)—,    -   (4) —CR^(2G)═CR^(2G)—,    -   (5) —S(═O)—,    -   (6) —SO₂—,    -   (7) —C(R^(3G))₂—S(═O)—,    -   (8) —S(═O)—C(R^(3G))₂—,    -   (9) —C(R^(3G))₂—SO₂—,    -   (10) —SO₂—C(R^(3G))₂—,    -   (11) —CR^(3G)═CR^(3G)—CR^(3G)═CR^(3G)—,    -   (12) —C(R^(3G))₂—,    -   (13) —CR^(3G)═CR^(3G)—C≡C—,    -   (14) —C≡C—CR^(3G)═CR^(3G)—,    -   (15) —C(═O)—C≡C—,    -   (16) —C≡C—C(═O)—,    -   (17) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (18) substituted or unsubstituted aryl,    -   (19) substituted or unsubstituted heterocyclyl, and    -   (20) substituted or unsubstituted heteroaryl,    -   wherein:        -   R^(1G) is substituted or unsubstituted C₁-C₆-alkyl;        -   each R^(2G) is independently selected from the group            consisting of H, a halogen atom, and substituted or            unsubstituted C₁-C₆-alkyl, and at least one R^(2G) is not H;            and        -   R^(3G) is selected from the group consisting of H, a halogen            atom, and substituted or unsubstituted C₁-C₆-alkyl;

Y is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

X is selected from the group consisting of:

-   -   (1) —(C═O)NR₄—,    -   (2) —C₁-C₆-alkyl-(C═O)NR₄—,    -   (3) —C₂-C₆-alkenyl-(C═O)NR₄—,    -   (4) —C₂-C₆-alkynyl-(C═O)NR₄—,    -   (5) —CH₂NR₄—,    -   (6) —SO₂NR₄—,    -   (7) —S(═O)NR₄—,    -   (8) —NR₄C(═O)—, and    -   (9) —NR₄—,    -   or X and A, together with the atoms to which they are attached        can form a heterocyclic ring, having from 5 to 8 ring atoms,        wherein 1-2 ring atoms of the heterocyclic ring are selected        from N, O and S,    -   or when Y is a bicyclic substituted or unsubstituted        heterocyclyl or heteroaryl, then X is absent;

R₃ is H or substituted or unsubstituted C₁-C₆-alkyl, or R₃ and A,together with the atom to which they are attached can form a substitutedor unsubstituted 3-10 membered cycloalkyl or a heterocyclic ring, havingfrom 3 to 10 ring atoms, wherein 1-4 ring atoms of the heterocyclic ringare selected from N, O and S;

R₄ is (1) H or substituted or unsubstituted C₁-C₆-alkyl, or (2) R₄ andA, together with the atoms to which they are attached can form asubstituted or unsubstituted heterocyclic ring, having from 3 to 8 ringatoms, wherein 1-2 ring atoms of the heterocyclic ring are selected fromN, O and S, or (3) R₄ and Y, together with the atoms to which they areattached, form a bicyclic substituted or unsubstituted heterocyclyl orheteroaryl;

n is an integer from 0-6;

A is selected from the group consisting of:

-   -   (1) H,    -   (2) —(CH₂)₀₋₄C(R^(1a),R^(2a))(CH₂)₀₋₄OR^(3a),    -   (3) —(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a),R^(5a)),    -   (4) —(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a))COR^(3a),    -   (5) —(CH₂)₀₋₄C(R^(1a),R^(2a))NHCON(R^(4a),R^(5a)),    -   (6) —(CH₂)₀₋₄C(R^(1a),R^(2a))NHC(═NH)N(R^(4a),R^(5a)),    -   (7) —CH(R^(1a),R^(2a)),    -   (8) —C≡CH,    -   (9) —(CH₂)₀₋₄C(R^(1a),R^(2a))CN,    -   (10) —(CH₂)₀₋₄C(R^(1a),R^(2a))CO₂R^(3a),    -   (11) —(CH₂)₀₋₄C(R^(1a),R^(2a))CON(R^(4a),R^(5a)),    -   (12) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (13) substituted or unsubstituted aryl,    -   (14) substituted or unsubstituted heterocyclyl, and    -   (15) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1a), R^(2a), R^(3a), R^(4a), and R^(5a) is            independently selected from the group consisting of:            -   (a) H,            -   (b) a halogen atom,            -   (c) substituted or unsubstituted C₁-C₆-alkyl,            -   (d) substituted or unsubstituted aryl,            -   (e) substituted or unsubstituted heterocyclyl, and            -   (f) substituted or unsubstituted heteroaryl,    -   or R^(4a) and R^(5a) together with the N atom to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 5 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S;

Q is absent or selected from the group consisting of:

-   -   (1) —C(═O)N(R₁,R₂),    -   (2) —NHC(═O)N(R₁,R₂),    -   (3) —N(OH)C(═O)N(R₁,R₂),    -   (4) —CH(OH)C(═O)N(R₁,R₂),    -   (5) —CH[N(R^(2q), R^(3q))]C(═O)N(R₁,R₂),    -   (6) —CHR¹⁹C(═O)N(R₁, R₂),    -   (7) —CO₂H,    -   (8) —C(═O)NHSO₂R^(4q),    -   (9) —SO₂NH₂,    -   (10) —N(OH)C(═O)R^(1q),    -   (11) —N(OH)SO₂R^(4q),    -   (12) —NHSO₂R^(4q),    -   (13) —SH,    -   (14) —CH(SH)(CH₂)₀₋₁C(═O)N(R₁,R₂),    -   (15) —CH(SH)(CH₂)₀₋₁CO₂H,    -   (16) —CH(OH)(CH₂)₀₋₁ CO₂H,    -   (17) —CH(SH)CH₂CO₂R^(1q),    -   (18) —CH(OH)(CH₂)SO₂NH₂,    -   (19) —CH(CH₂SH)NHCOR^(1q),    -   (20) —CH(CH₂SH)NHSO₂R^(4q),    -   (21) —CH(CH₂SR^(5q))CO₂H,    -   (22) —CH(CH₂SH)NHSO₂NH₂,    -   (23) —CH(CH₂OH)CO₂H,    -   (24) —CH(CH₂OH)NHSO₂NH₂,    -   (25) —C(═O)CH₂CO₂H,    -   (26) —C(═O)(CH₂)₀₋₁CONH₂,    -   (27) —OSO₂NHR^(5q),    -   (28) —SO₂NHNH₂,    -   (29) —P(═O)(OH)₂,

-   -   and    -   (33) —N(OH)C(═O)CR₁R₂,    -   wherein:        -   R₁ is selected from the group consisting of:            -   (1) —H,            -   (2) —OH,            -   (3) —OC₁-C₆-alkyl,            -   (4) —N(R^(2q), R^(3q)), and            -   (5) substituted or unsubstituted C₁-C₆-alkyl; R₂ is                selected from the group consisting of:        -   (1) H,            -   (2) substituted or unsubstituted C₁-C₆-alkyl,            -   (3) substituted or unsubstituted C₂-C₆-alkenyl,            -   (4) substituted or unsubstituted C₂-C₆-alkenyl,            -   (5) substituted or unsubstituted aryl,            -   (6) substituted or unsubstituted heterocyclyl, and            -   (7) substituted or unsubstituted heteroaryl,        -   or R₁ and R₂, together with the N atom to which they are            attached can form a substituted or unsubstituted            heterocyclic ring, having from 3 to 10 ring atoms, wherein            1-4 ring atoms of the heterocyclic ring are selected from N,            O and S; and        -   each R^(1q), R^(2q), R^(3q), R^(4q), and R^(5q) is            independently selected from the group consisting of H and            C₁-C₆ alkyl.

In certain embodiments of the above embodiment, G is selected from thegroup consisting of:

-   -   (1) —C≡C—,    -   (2) —C≡C—C≡C—,    -   (3) —CR^(3G)═CR^(3G)—C≡C—, and    -   (4) —C≡C—CR^(3G)═CR^(3G)—.        For example, G may be —C≡C—, —C≡C—C≡C—, —CH═CH—C≡C— or        —C≡C—CH═CH—. In embodiments wherein G is —CH═CH—C≡C—, G may have        one of the following structures:

and in embodiments wherein G is —C≡C—CH═CH—, G may have one of thefollowing structures:

In certain embodiments of the above embodiments, X is —(C═O)NR₄—. Forexample, X may be —(C═O)NH—.

In certain embodiments of the above embodiments, Q is —(C═O)N(R₁,R₂).For example, Q may be —(C═O)NHOH.

In certain embodiments of the above embodiments, n is 0.

In certain embodiments of the above embodiments, R₃ is H.

In certain embodiments of the above embodiments, Y is substituted orunsubstituted aryl. For example, Y may be substituted or unsubstitutedphenyl.

In certain embodiments of the above embodiments, A is selected from thegroup consisting of:

-   -   (1) —(CH₂)₀₋₄C(R^(1a),R^(2a))(CH₂)₀₋₄OR^(3a),    -   (2) —(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a),R^(5a)), and    -   (3) —CH(R^(1a),R^(2a)).        For example, A may be —CH(CH₃)₂, —CH₂OH, —CH₂NH₂, —CHCH₃OH,        —CHCH₃NH₂ or —C(CH₃)₂OH, or A may be —C(CH₃)₂NH₂.

In other certain embodiments of the above embodiments, A is selectedfrom the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl.        For example, A may be:

In certain embodiments of the above embodiments, R₃ is H and

has the following structure:

For example, when A is —CHCH₃OH or —CHCH₃NH₂,

has one of the following structures:

In certain embodiments of the above embodiments, D is present. Forexample, D may be substituted or unsubstituted heteroaryl (such as, forexample, a heteroaryl is selected from the group consisting of:

Alternatively, D may be substituted or unsubstituted aryl (such as, forexample, substituted or unsubstituted phenyl),

In other certain embodiments of the above embodiments, D is absent.

In certain embodiments of the above embodiments, L is present. Forexample, L may be substituted or unsubstituted alkyl (such as —CH₃—) orL may be —CH₂—NH—.

In other certain embodiments of the above embodiments, L is absent.

In a second embodiment, the present invention provides compounds offormula (I) wherein:

E is selected from the group consisting of:

-   -   (1) H,    -   (2) substituted or unsubstituted C₁-C₆-alkyl,    -   (3) substituted or unsubstituted C₂-C₆-alkenyl,    -   (4) substituted or unsubstituted C₂-C₆-alkynyl,    -   (5) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (6) substituted or unsubstituted aryl,    -   (7) substituted or unsubstituted heterocyclyl, and    -   (8) substituted or unsubstituted heteroaryl;

L is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₁-C₆-alkyl,    -   (2) —(NR^(3L))₀₋₁—(CH₂)₀₋₄—NR^(3L)—(CH₂)₀₋₄—,    -   (3) —(NR^(3L))₀₋₁—C(R^(1L),R^(2L))—NR^(3L) C(R^(1L),R^(2L))—,    -   (4) —C(R^(1L),R^(2L))—O—C(R^(1L),R^(2L))—,    -   (5) —(CH₂)₀₋₄—NR^(3L)—C(R^(1L),R^(2L))—CONH—(CH₂)₀₋₄—,    -   (6) —CO—C(R^(1L),R^(2L))—NHCO—,    -   (7) —CONR^(3L)—,    -   (8) —NR^(3L)CO—,    -   (9) —NR^(3L)—,    -   (10) —SO₂NR^(3L)—,    -   (11) —NR^(3L)—C(═O)—NR^(3L)—,    -   (12) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (13) substituted or unsubstituted aryl,    -   (14) substituted or unsubstituted heterocyclyl, and    -   (15) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1L), R^(2L), and R^(3L) is independently selected            from the group consisting of:            -   (a) H,            -   (b) substituted or unsubstituted C₁-C₆-alkyl,            -   (c) C₁-C₆-alkyl substituted with aryl,            -   (d) C₁-C₆-alkyl substituted with heterocyclyl, and            -   (e) C₁-C₆-alkyl substituted with heteroaryl,    -   or R^(1L) and R^(3L), together with the atoms to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 3 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S;

D is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

G is selected from the group consisting of:

-   -   (1) —(CH₂)₀₋₄—O—(CH₂)₀₋₄—,    -   (2) —(CH₂)₀₋₄—S—(CH₂)₀₋₄—,    -   (3) —(CH₂)₀₋₄—NR^(1G)—(CH₂)₀₋₄—,    -   (4) —C(═O)—,    -   (5) —NR^(1G)C(═O)—,    -   (6) —C(═O)NR^(1G)—,    -   (7) —(CH₂)₀₋₄NHCH₂C(═O)NR^(1G)—,    -   (8) —C≡C—,    -   (9) —C≡C—C≡C—,    -   (10) —CR^(2G)═CR^(2G)—,    -   (11) —S(═O)—,    -   (12) —SO₂—,    -   (13) —C(R^(3G))₂—S(═O)—,    -   (14) —S(═O)—C(R^(3G))₂—,    -   (15) —C(R^(3G))₂—SO₂—,    -   (16) —SO₂—C(R^(3G))₂—,    -   (17) —CR^(3G)═CR^(3G)—CR^(3G)═CR^(3G)—,    -   (18) —C(R^(3G))₂—,    -   (19) —CR^(3G)═CR^(3G)—C≡C—,    -   (20) —C≡C—CR^(3G)═CR^(3G)—,    -   (21) —C(═O)—C≡C—,    -   (22) —C≡C—C(═O)—,    -   (23) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (24) substituted or unsubstituted aryl,    -   (25) substituted or unsubstituted heterocyclyl, and    -   (26) substituted or unsubstituted heteroaryl,    -   wherein:        -   R^(1G) is substituted or unsubstituted C₁-C₆-alkyl;        -   each R^(2G) and R^(3G) is independently selected from the            group consisting of H, a halogen atom, and substituted or            unsubstituted C₁-C₆-alkyl;

Y is absent or selected from the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl;

X is selected from the group consisting of:

-   -   (1) —(C═O)NR₄—,    -   (2) —C₁-C₆-alkyl-(C═O)NR₄—,    -   (3) —C₂-C₆-alkenyl-(C═O)NR₄—,    -   (4) —C₂-C₆-alkynyl-(C═O)NR₄—,    -   (5) —CH₂NR₄—,    -   (6) —SO₂NR₄—,    -   (7) —S(═O)NR₄—,    -   (8) —NR₄C(═O)—, and    -   (9) —NR₄—,    -   or X and A, together with the atoms to which they are attached        can form a heterocyclic ring, having from 5 to 8 ring atoms,        wherein 1-2 ring atoms of the heterocyclic ring are selected        from N, O and S,    -   or when Y is a bicyclic substituted or unsubstituted        heterocyclyl or heteroaryl, then X is absent;

R₃ is H or substituted or unsubstituted C₁-C₆-alkyl, or R₃ and A,together with the atom to which they are attached can form a substitutedor unsubstituted 3-10 membered cycloalkyl or a heterocyclic ring, havingfrom 3 to 10 ring atoms, wherein 1-4 ring atoms of the heterocyclic ringare selected from N, O and S;

R₄ is (1) H or substituted or unsubstituted C₁-C₆-alkyl, or (2) R₄ andA, together with the atoms to which they are attached can form asubstituted or unsubstituted heterocyclic ring, having from 3 to 8 ringatoms, wherein 1-2 ring atoms of the heterocyclic ring are selected fromN, O and S, or (3) R₄ and Y, together with the atoms to which they areattached, form a bicyclic substituted or unsubstituted heterocyclyl orheteroaryl;

n is an integer from 0-6;

A is selected from the group consisting of:

-   -   (1) —C(R^(1a),R^(2a))OR^(3a),    -   (2) —C(R^(1a),R^(2a))N(R^(4a),R^(5a)),    -   (3) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (4) substituted or unsubstituted aryl,    -   (5) substituted or unsubstituted heterocyclyl, and    -   (6) substituted or unsubstituted heteroaryl,    -   wherein:        -   each R^(1a) and R^(2a) is independently selected from the            group consisting of substituted or unsubstituted            C₁-C₆-alkyl;        -   each R^(3a), R^(4a), and R^(5a) is independently selected            from the group consisting of:            -   (a) H,            -   (b) a halogen atom,            -   (c) substituted or unsubstituted C₁-C₆-alkyl,            -   (d) substituted or unsubstituted aryl,            -   (e) substituted or unsubstituted heterocyclyl, and            -   (f) substituted or unsubstituted heteroaryl,    -   or R^(4a) and R^(5a) together with the N atom to which they are        attached can form a substituted or unsubstituted heterocyclic        ring, having from 5 to 8 ring atoms, wherein 1-2 ring atoms of        the heterocyclic ring are selected from N, O and S; and        -   when A is —C(R^(1a),R^(2a))OR^(3a), the compound is not            2-{[(4′-ethyl-1,1′-biphenyl-4-yl)carbonyl]amino}-3-hydroxy-3-methylbutanoic            acid, 4′-ethyl-N-{2-hydroxy-1-[(hydroxyamino)            carbonyl]-2-methylpropyl}-1,1′-biphenyl-4-carboxamide or            N-{2-hydroxy-1-[(hydroxyamino)carbonyl]-2-methylpropyl}-4-(phenylethynyl)benzamide;

Q is absent or selected from the group consisting of:

-   -   (1) —C(═O)N(R₁,R₂),    -   (2) —NHC(═O)N(R₁,R₂),    -   (3) —N(OH)C(═O)N(R₁,R₂),    -   (4) —CH(OH)C(═O)N(R₁,R₂),    -   (5) —CH[N(R^(2q), R^(3q))]C(═O)N(R₁,R₂),    -   (6) —CHR^(1q)C(═O)N(R₁, R₂),    -   (7) —CO₂H,    -   (8) —C(═O)NHSO₂R^(4q),    -   (9) —SO₂NH₂,    -   (10) —N(OH)C(═O)R^(1q),    -   (11) —N(OH)SO₂R^(4q),    -   (12) —NHSO₂R^(4q),    -   (13) —SH,    -   (14) —CH(SH)(CH₂)₀₋₁C(═O)N(R₁,R₂),    -   (15) —CH(SH)(CH₂)₀₋₁CO₂H,    -   (16) —CH(OH)(CH₂)₀₋₁CO₂H,    -   (17) —CH(SH)CH₂CO₂R^(1q),    -   (18) —CH(OH)(CH₂)SO₂NH₂,    -   (19) —CH(CH₂SH)NHCOR^(1q),    -   (20) —CH(CH₂SH)NHSO₂R^(4q),    -   (21) —CH(CH₂SR^(5q))CO₂H,    -   (22) —CH(CH₂SH)NHSO₂NH₂,    -   (23) —CH(CH₂OH)CO₂H,    -   (24) —CH(CH₂OH)NHSO₂NH₂,    -   (25) —C(═O)CH₂CO₂H,    -   (26) —C(═O)(CH₂)₀₋₁CONH₂,    -   (27) —OSO₂NHR^(5q),    -   (28) —SO₂NHNH₂,    -   (29) —P(═O)(OH)₂,

-   -   and    -   (33) —N(OH)C(═O)CR₁R₂,    -   wherein:        -   R₁ is selected from the group consisting of:            -   (1) —H,            -   (2) —OH,            -   (3) —OC₁-C₆-alkyl,            -   (4) —N(R^(2q), R^(3q)), and            -   (5) substituted or unsubstituted C₁-C₆-alkyl;        -   R₂ is selected from the group consisting of:            -   (1) H,            -   (2) substituted or unsubstituted C₁-C₆-alkyl,            -   (3) substituted or unsubstituted C₂-C₆-alkenyl,            -   (4) substituted or unsubstituted C₂-C₆-alkenyl,            -   (5) substituted or unsubstituted aryl,            -   (6) substituted or unsubstituted heterocyclyl, and            -   (7) substituted or unsubstituted heteroaryl,        -   or R₁ and R₂, together with the N atom to which they are            attached can form a substituted or unsubstituted            heterocyclic ring, having from 3 to 10 ring atoms, wherein            1-4 ring atoms of the heterocyclic ring are selected from N,            O and S; and        -   each R^(1q), R^(2q), R^(3q), R^(4q), and R^(5q) is            independently selected from the group consisting of H and            C₁-C₆ alkyl.

In certain embodiments of the above embodiment, A is—C(R^(1a),R^(2a))N(R^(4a),R^(5a)). For example, A may be —C(CH₃)₂NH₂.

In other certain embodiments of the above embodiment, A is—C(R^(1a),R^(2a))OR^(3a).

In other certain embodiments of the above embodiments, A is selectedfrom the group consisting of:

-   -   (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl,    -   (2) substituted or unsubstituted aryl,    -   (3) substituted or unsubstituted heterocyclyl, and    -   (4) substituted or unsubstituted heteroaryl.        For example, A may be:

In certain embodiments of the above embodiments, R₃ is H and

has the following structure:

In certain embodiments of the above embodiments, G is selected from thegroup consisting of:

-   -   (1) —C≡C—,    -   (2) —C≡C—C≡C—,    -   (3) —CR^(3G)═CR^(3G)—C≡C—, and    -   (4) —C≡C—CR^(3G)═CR^(3G)—.        For example, G may be —C≡C—, —C≡C—C≡C—, —CH═CH—C≡C—, or        —C≡C—CH═CH—. In embodiments wherein G is —CH═CH—C≡C—, G may have        one of the following structures:

and in embodiments wherein G is —C≡C—CH═CH—, G may have one of thefollowing structures:

In certain embodiments of the above embodiments, X is —(C═O)NR₄—. Forexample, X may be —(C═O)NH—.

In certain embodiments of the above embodiments, Q is —(C═O)N(R₁,R₂).For example, Q may be —(C═O)NHOH.

In certain embodiments of the above embodiments, n is 0.

In certain embodiments of the above embodiments, R₃ is H.

In certain embodiments of the above embodiments, Y is substituted orunsubstituted aryl. For example, Y may be substituted or unsubstitutedphenyl.

In certain embodiments of the above embodiments, D is present. Forexample, D may be substituted or unsubstituted heteroaryl (such as, forexample, a heteroaryl is selected from the group consisting of:

Alternatively, D may be substituted or unsubstituted aryl (such as, forexample, substituted or unsubstituted phenyl),

In other certain embodiments of the above embodiments, D is absent.

In certain embodiments of the above embodiments, L is present. Forexample, L may be substituted or unsubstituted alkyl (such as —CH₃—) orL may be —CH₂—NH—.

In other certain embodiments of the above embodiments, L is absent.

In various other embodiments of all of the foregoing embodiments,E-L-D-G-Y taken together, may be selected from the group consisting of:

wherein R is selected from the group consisting of —CH₃, —C₂H₅, —CH₂OH,—OH, —OCH₃, —OC₂H₅, —OCF₃, —CN, —NO₂, —CO₂, —CO₂CH₃, —CONH₂, —NH₂, —F,—Cl, —Br, —CF₃, —N(CH₃)₂, —NHSO₂CH₃, and —NHCOCH₃.

In various other embodiments of all of the foregoing embodiments, G maybe selected from the group consisting of:

In various other embodiments of all of the foregoing embodiments, A maybe:

In various other embodiments of all of the foregoing embodiments, thecompound may have the structure:

In various other embodiments of all of the foregoing embodiments, X is—(C═O)NR₄—, and R₄ and A, together with the atoms to which they areattached, form a substituted heterocyclic ring, and the compounds havethe following structure:

In various other embodiments of all of the foregoing embodiments, X is—(C═O)NR₄—, and the compounds have the following structure:

In various other embodiments of all of the foregoing embodiments, X andA, together with the atoms to which they are attached form aheterocyclic ring, and the compounds have the following structure:

In various other embodiments of all of the foregoing embodiments, Q is—N(OH)C(═O)CR₁R₂ and at least one of R₁ and R₂ is not H or C₁-C₆-alkyl.

In various other embodiments of all of the foregoing embodiments, X isselected from the group consisting of:

-   -   (1) —SO₂NR₄—,    -   (2) —S(═O)NR₄—,    -   (3) —NR₄C(═O)—, and    -   (4) —NR₄—,        wherein R₄ is H or substituted or unsubstituted C₁-C₆-alkyl, or        R₄ and A, together with the atoms to which they are attached can        form a substituted or unsubstituted heterocyclic ring, having        from 3 to 8 ring atoms, wherein 1-2 ring atoms of the        heterocyclic ring are selected from N, O and S,

or X is selected from the group consisting of:

-   -   (1) —(C═O)NR₄—,    -   (2) —C₁-C₆-alkyl-(C═O)NR₄—,    -   (3) —C₂-C₆-alkenyl-(C═O)NR₄—,    -   (4) —C₂-C₆-alkynyl-(C═O)NR₄—,    -   (5) —CH₂NR₄—,    -   (6) —SO₂NR₄—,    -   (7) —S(═O)NR₄—,    -   (8) —NR₄C(═O)—, and    -   (9) —NR₄—,        wherein R₄ and Y, together with the atoms to which they are        attached, form a bicyclic substituted or unsubstituted        heterocyclyl or heteroaryl,

or when Y is a bicyclic substituted or unsubstituted heterocyclyl orheteroaryl, then X is absent.

For example, in more specific embodiments, X may be —NH— and Y may be abicyclic substituted or unsubstituted heterocyclyl or heteroaryl. Insuch embodiments, Y may be selected from the group consisting of:

wherein: R^(y1) is S, O, NH or N(C₁-C₆-alkyl); R^(y2) is N, CH, orC(C₁-C₆-alkyl); and R^(y3) is S or 0.

In other more specific embodiments, X may be —(C═O)NR₄— and R₄ and Y,together with the atoms to which they are attached, may form a bicyclicsubstituted or unsubstituted heterocyclyl or heteroaryl. In suchembodiments, R₄ and Y may form:

In other more specific embodiments, Y may be a bicyclic substituted orunsubstituted heterocyclyl or heteroaryl and X may be absent. In suchembodiments, Y may be:

In various other embodiments of all of the foregoing embodiments,—X—C(AR₃)—(CH₂)_(n)-Q is —X₁-Q₁ wherein:

X₁ is absent or selected from the group consisting of:

-   -   (1) —(C═O)—,    -   (2) —CR^(x)—,    -   (3) —(C═O)NR^(x)—, and    -   (4) —NR^(x)(C═O)—,    -   wherein R^(x) is H or substituted or unsubstituted C₁-C₆-alkyl;

Q₁ is selected from the group consisting of:

-   -   wherein:        -   R^(1Q) is a carbon or a nitrogen ring atom,        -   each R^(2Q) is absent or is a carbon ring atom,        -   R^(1Q) and an adjacent R^(2Q) optionally form a double bond,            and        -   each carbon ring atom is optionally substituted with a            substituted or unsubstituted C₁-C₆ alkyl, or an oxo            substituent.

For example, —X₁-Q₁ may be:

wherein R^(3Q) is H or substituted or unsubstituted C₁-C₆ alkyl.

In various other embodiments or all of the foregoing embodiments, Acontains at least one halogen atom.

In another aspect, the invention provides a method of inhibiting adeacetylase enzyme in a gram-negative bacteria, thereby affectingbacterial growth, comprising administering to a patient in need of suchinhibition a compound of formula (I).

In another aspect, the invention provides a method of inhibiting LpxC,thereby modulating the virulence of a bacterial infection, comprisingadministering to a patient in need of such inhibition a compound offormula (I). In certain embodiments of the method of inhibiting LpxCusing a compound of formula (I), the IC₅₀ value of the compound is lessthan or equal to 10 μM with respect to LpxC. In other embodiments, theIC₅₀ value is less than or equal to 1 μM, is less than or equal to 0.1μM, is less than or equal to 0.050 μM, is less than or equal to 0.030μM, is less than or equal to 0.025 μM, or is less than or equal to 0.010μM.

In another aspect, the invention provides a method for treating asubject with a gram-negative bacterial infection comprisingadministering to the subject in need thereof an antibacteriallyeffective amount of a compound of formula (I) with a pharmaceuticallyacceptable carrier. In certain embodiments, the subject may be a mammal,and in some embodiments, a human.

In another aspect, the invention provides a method of administering aninhibitory amount of a compound of formula (I) to fermentative ornon-fermentative gram-negative bacteria. In a more specific embodimentof the method of administering an inhibitory amount of a compound offormula (I) to fermentative or non-fermentative gram-negative bacteria,the gram-negative bacteria are selected from the group consisting ofPseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderiacepacia, Alcaligenes xylosoxidans, Acinetobacter, Enterobacteriaceae,Haemophilus, Franciscellaceae (Franciscella tularensis) and Neisseriaspecies.

In another aspect, the invention provides a method of administering aninhibitory amount of a compound of formula (I) to gram-negativebacteria, such as Enterobacteriaceae which is selected from the groupconsisting of organisms such as Serratia, Proteus, Klebsiella,Enterobacter, Citrobacter, Salmonella, Providencia, Yersinia (Yersiniapestis), Morganella, Cedecea, and Edwardsiella species and Escherichiacoli.

In another aspect, the invention provides a pharmaceutical compositionor formulation comprising an effective amount of a compound of formula(I) with a pharmaceutically acceptable carrier thereof.

In another aspect, the invention provides a method of co-administering acompound of formula (I) with other therapeutic agents that are selectedfor their particular usefulness against the condition that is beingtreated. For example, a compound of formula (I) is useful in combinationwith other anti-bacterial agents. The compound of formula (I) augmentsthe sensitivity of gram-negative bacteria to existing classes ofantibacterials. Combinations of the presently disclosed compounds withother anti-bacterial agents are within the scope of the invention. Suchanti-bacterial agents include, but are not limited to, erythromycin,rifampicin, Nalidixic acid, carbenicillin, bacitracin, cycloserine,fosfomycin, and vancomycin.

A further aspect of the invention is the use of LpxC inhibitors for thetreatment of an infection, particularly a bacterial infection. Abacterial infection treated with the compounds of the invention can be aprimary infection or a co-infection caused by a species of bacteria andone or more additional infectious agents selected from the groupconsisting of bacteria, virus, parasite and fungus.

The term “treating”, as used herein, refers to reversing, alleviating,inhibiting the progress of, or preventing the disorder or condition towhich such term applies, or one or more symptoms of such disorder orcondition. The term “treatment”, as used herein, refers to the act oftreating, as “treating” is defined immediately above.

The compounds of the invention can be used for treating conditionscaused by the bacterial production of endotoxin and, in particular, bygram-negative bacteria and bacteria that use LpxC in the biosynthesis oflipopolysaccharide (LPS) or endotoxin.

The compounds of the invention also are useful in the conditions thatare caused or exacerbated by the bacterial production of lipid A and LPSor endotoxin, such as sepsis, septic shock, systemic inflammation,localized inflammation, chronic obstructive pulmonary disease (COPD) andacute exacerbations of chronic bronchitis (AECB). For these conditions,treatment includes the administration of a compound of the invention, ora combination of compounds of the invention, optionally with a secondagent wherein the second agent is a second antibacterial agent or asecond non-antibacterial agent.

For sepsis, septic shock, systemic inflammation, localized inflammation,chronic obstructive pulmonary disease (COPD) and acute exacerbations ofchronic bronchitis (AECB), preferred second non-antibacterial agentsinclude antiendotoxins including endotoxin receptor-binding antibodies,endotoxin-binding antibodies, anti-CD14-binding protein antibodies,antilipopolysaccharide-binding protein antibodies and tyrosine kinaseinhibitors.

In treatment of serious or chronic respiratory tract infections, thecompounds of the present invention may also be used with secondnon-antibacterial agents administered via inhalation. Representativenon-antibacterial agents used in this treatment includeanti-inflammatory steroids, non-steroidal anti-inflammatory agents,bronchiodilators, mucolytics, anti-asthma therapeutics and lung fluidsurfactants. In particular, the non-antibacterial agent may be selectedfrom a group consisting of albuterol, salbuterol, budesonide,beclomethasone, dexamethasone, nedocromil, beclomethasone, fluticasone,flunisolide, triamcinolone, ibuprofin, rofecoxib, naproxen, celecoxib,nedocromil, ipratropium, metaproterenol, pirbuterol, salmeterol,formoterol, indacaterol, bronchiodilators, mucolytics, calfactant,beractant, poractant alfa, surfaxin and pulmozyme (also called domasealfa).

The compounds of the invention can be used, alone or in combination witha second antibacterial agent for the treatment of a serious or chronicrespiratory tract infection including serious lung and nosocomialinfections such as those caused by Enterobacter aerogenes, Enterobactercloacae, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca,Proteus mirabilis, Serratia marcescens, Stenotrophomonas maltophilia,Pseudomonas aeruginosa, Burkholderia cepacia, Acinetobactercalcoaceticus, Alcaligenes xylosoxidans, Flavobacterium meningosepticum,Providencia stuartii and Citrobacter freundi, community lung infectionssuch as those caused by Haemophilus Influenzae, Legionella species,Moraxella catarrhalis, Branhamella catarrhalis, Enterobacter species,Acinetobacter species, Klebsiella species, and Proteus species, andinfections caused by other bacterial species such as Neisseria species,Shigella species, Salmonella species, Helicobacter pylori, Vibrionaceaeand Bordetella species as well as the infections is caused by a Brucellaspecies, Francisella tularensis and/or Yersinia Pestis.

When used for treating subjects infected with gram-negative bacterialinfections, the compounds of the present invention can be used tosensitize gram-negative bacteria to the effects of a second agent.

When the compounds of the present invention are used in combination witha second antibacterial agent, non-limiting examples of antibacterialagents may be selected from the following groups:

(1) Macrolides or ketolides such as erythromycin, azithromycin,clarithromycin and telithromycin;

(2) Beta-lactams such as penicillin G, penicillin V, methicillin,oxacillin, cloxacillin, dicloxacillin, nafcillin, ampicillin,amoxicillin, carbenicillin, ticarcillin, mezlocillin, piperacillin,azlocillin, temocillin, cepalothin, cephapirin, cephradine,cephaloridine, cefazolin, cefamandole, cefuroxime, cephalexin,cefprozil, cefaclor, loracarbef, cefoxitin, cefinetazole, cefotaxime,ceftizoxime, ceftriaxone, cefoperazone, ceftazidime, cefixime,cefpodoxime, ceftibuten, cefdinir, cefpirome, cefepime, aztreonam,imipenem, meropenem, ertapenem, doripenem, ceftobiprole, andceftaroline;

(3) Quinolones such as nalidixic acid, oxolinic acid, norfloxacin,pefloxacin, enoxacin, ofloxacin, levofloxacin, ciprofloxacin,temafloxacin, lomefloxacin, fleroxacin, grepafloxacin, sparfloxacin,trovafloxacin, clinafloxacin, gatifloxacin, moxifloxacin, sitafloxacin,garenoxacin, gemifloxacin and pazufloxacin;

(4) Antibacterial sulfonanmides and antibacterial sulphanilamides,including para-aminobenzoic acid, sulfadiazine, sulfisoxazole,sulfamethoxazole and sulfathalidine;

(5) Aminoglycosides such as streptomycin, neomycin, kanamycin,paromycin, gentamicin, tobramycin, amikacin, netilmicin, spectinomycin,sisomicin, dibekacin and isepamicin;

(6) Tetracyclines such as tetracycline, chlortetracycline,demeclocycline, minocycline, oxytetracycline, methacycline, tigecycline,doxycycline;

(7) Rifamycins such as rifampicin (also called rifampin), rifapentine,rifabutin, bezoxazinorifamycin and rifaximin;

(8) Lincosamides such as lincomycin and clindamycin;

(9) Glycopeptides such as telavancin, vancomycin and teicoplanin orlipopeptides such as daptomycin;

(10) Streptogramins such as quinupristin and daflopristin;

(11) Oxazolidinones such as linezolid;

(12) Polymyxin, colistin and colymycin; and

(13) Trimethoprim and bacitracin.

The second antibacterial agent may be administered in combination withthe compounds of the present inventions, wherein the secondantibacterial agent is administered prior to, simultaneously, or afterthe compound or compounds of the present invention. When simultaneousadministration of a compound of the invention with a second agent isdesired and the route of administration is the same, then a compound ofthe invention may be formulated with a second agent into the same dosageform. An example of a dosage form containing a compound of the inventionand a second agent is a tablet or a capsule.

When used for treating a serious or chronic respiratory tractinfections, the compounds of the invention may be used alone or incombination with a second antibacterial agent administered viainhalation. In the case of inhalation, a preferred second antibacterialagent is selected from a group consisting of tobramycin, gentamicin,aztreonam, ciprofloxacin, polymyxin, colistin, colymycin, azithromycinand clarithromycin.

Pharmaceutical Compositions

Pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials that can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), bucally, or as an oral or nasal spray, or a liquid aerosol ordry powder formulation for inhalation.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions that can bedissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionthat, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform may be accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations may also be prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories that can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,acetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulations, ear drops, and the like are also contemplatedas being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Compositions of the invention may also be formulated for delivery as aliquid aerosol or inhalable dry powder. Liquid aerosol formulations maybe nebulized predominantly into particle sizes that can be delivered tothe terminal and respiratory bronchioles where bacteria reside inpatients with bronchial infections, such as chronic bronchitis andpneumonia. Pathogenic bacteria are commonly present throughout airwaysdown to bronchi, bronchioli and lung parenchema, particularly interminal and respiratory bronchioles. During exacerbation of infection,bacteria can also be present in alveoli. Liquid aerosol and inhalabledry powder formulations are preferably delivered throughout theendobronchial tree to the terminal bronchioles and eventually to theparenchymal tissue.

Aerosolized formulations of the invention may be delivered using anaerosol forming device, such as a jet, vibrating porous plate orultrasonic nebulizer, preferably selected to allow the formation of aaerosol particles having with a mass medium average diameterpredominantly between 1 to 5 μm. Further, the formulation preferably hasbalanced osmolarity ionic strength and chloride concentration, and thesmallest aerosolizable volume able to deliver effective dose of thecompounds of the invention to the site of the infection. Additionally,the aerosolized formulation preferably does not impair negatively thefunctionality of the airways and does not cause undesirable sideeffects.

Aerosolization devices suitable for administration of aerosolformulations of the invention include, for example, jet, vibratingporous plate, ultrasonic nebulizers and energized dry powder inhalers,that are able to nebulize the formulation of the invention into aerosolparticle size predominantly in the size range from 1-5 pm. Predominantlyin this application means that at least 70% but preferably more than 90%of all generated aerosol particles are 1 to 5 μm range. A jet nebulizerworks by air pressure to break a liquid solution into aerosol droplets.Vibrating porous plate nebulizers work by using a sonic vacuum producedby a rapidly vibrating porous plate to extrude a solvent droplet througha porous plate. An ultrasonic nebulizer works by a piezoelectric crystalthat shears a liquid into small aerosol droplets. A variety of suitabledevices are available, including, for example, AeroNeb and AeroDosevibrating porous plate nebulizers (AeroGen, Inc., Sunnyvale, Calif.),Sidestream7 nebulizers (Medic-Aid Ltd., West Sussex, England), Pari LC7and Pari LC Star7 jet nebulizers (Pari Respiratory Equipment, Inc.,Richmond, Va.), and Aerosonic (DeVilbiss Medizinische Produkte(Deutschland) GmbH, Heiden, Germany) and UltraAire7 (Omron Healthcare,Inc., Vernon Hills, Ill.) ultrasonic nebulizers.

Compounds of the invention may also be formulated for use as topicalpowders and sprays that can contain, in addition to the compounds ofthis invention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

According to the methods of treatment of the present invention,bacterial infections are treated or prevented in a patient such as ahuman or lower mammal by administering to the patient a therapeuticallyeffective amount of a compound of the invention, in such amounts and forsuch time as is necessary to achieve the desired result. By a“therapeutically effective amount” of a compound of the invention ismeant a sufficient amount of the compound to treat bacterial infections,at a reasonable benefit/risk ratio applicable to any medical treatment.It will be understood, however, that the total daily usage of thecompounds and compositions of the present invention will be decided bythe attending physician within the scope of sound medical judgment. Thespecific therapeutically effective dose level for any particular patientwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed; andlike factors well known in the medical arts.

The total daily dose of the compounds of this invention administered toa human or other mammal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 2000 mg of the compound(s) of this invention per day in singleor multiple doses.

Methods of formulation are well known in the art and are disclosed, forexample, in Remington: The Science and Practice of Pharmacy, MackPublishing Company, Easton, Pa., 19th Edition (1995). Pharmaceuticalcompositions for use in the present invention can be in the form ofsterile, non-pyrogenic liquid solutions or suspensions, coated capsules,suppositories, lyophilized powders, transdermal patches or other formsknown in the art.

A “kit” as used in the instant application includes a container forcontaining the pharmaceutical compositions and may also include dividedcontainers such as a divided bottle or a divided foil packet. Thecontainer can be in any conventional shape or form as known in the artthat is made of a pharmaceutically acceptable material, for example apaper or cardboard box, a glass or plastic bottle or jar, a resealablebag (for example, to hold a “refill” of tablets for placement into adifferent container), or a blister pack with individual doses forpressing out of the pack according to a therapeutic schedule. Thecontainer employed can depend on the exact dosage form involved, forexample a conventional cardboard box would not generally be used to holda liquid suspension. It is feasible that more than one container can beused together in a single package to market a single dosage form. Forexample, tablets may be contained in a bottle that is in turn containedwithin a box.

An example of such a kit is a so-called blister pack. Blister packs arewell known in the packaging industry and are being widely used for thepackaging of pharmaceutical unit dosage forms (tablets, capsules, andthe like). Blister packs generally consist of a sheet of relativelystiff material covered with a foil of a preferably transparent plasticmaterial. During the packaging process, recesses are formed in theplastic foil. The recesses have the size and shape of individual tabletsor capsules to be packed or may have the size and shape to accommodatemultiple tablets and/or capsules to be packed. Next, the tablets orcapsules are placed in the recesses accordingly and the sheet ofrelatively stiff material is sealed against the plastic foil at the faceof the foil that is opposite from the direction in which the recesseswere formed. As a result, the tablets or capsules are individuallysealed or collectively sealed, as desired, in the recesses between theplastic foil and the sheet. Preferably the strength of the sheet is suchthat the tablets or capsules can be removed from the blister pack bymanually applying pressure on the recesses whereby an opening is formedin the sheet at the place of the recess. The tablet or capsule can thenbe removed via said opening.

It maybe desirable to provide a written memory aid, where the writtenmemory aid is of the type containing information and/or instructions forthe physician, pharmacist or other health care provider, or subject,e.g., in the form of numbers next to the tablets or capsules whereby thenumbers correspond with the days of the regimen that the tablets orcapsules so specified should be ingested or a card that contains thesame type of information. Another example of such a memory aid is acalendar printed on the card e.g., as follows “First Week, Monday,Tuesday,” . . . etc. . . . “Second Week, Monday, Tuesday, . . . ” etc.Other variations of memory aids will be readily apparent. A “daily dose”can be a single tablet or capsule or several tablets or capsules to betaken on a given day. When the kit contains separate compositions, adaily dose of one or more compositions of the kit can consist of onetablet or capsule while a daily dose of another one or more compositionsof the kit can consist of several tablets or capsules.

Another specific embodiment of a kit is a dispenser designed to dispensethe daily doses one at a time in the order of their intended use.Preferably, the dispenser is equipped with a memory-aid, so as tofurther facilitate compliance with the regimen. An example of such amemory-aid is a mechanical counter, that indicates the number of dailydoses that has been dispensed. Another example of such a memory-aid is abattery-powered micro-chip memory coupled with a liquid crystal readout,or audible reminder signal that, for example, reads out the date thatthe last daily dose has been taken and/or reminds one when the next doseis to be taken.

The kits of the present invention may also include, in addition to LpxCinhibitors, one or more additional pharmaceutically active compounds.Preferably, the additional compound is another LpxC inhibitor or anothercompound useful to bacterial infections. The additional compounds may beadministered in the same dosage form as the LpxC inhibitor or indifferent dosage forms. Likewise, the additional compounds can beadministered at the same time as the LpxC inhibitor or at differenttimes.

Compositions of the present compounds may also be used in combinationwith other known antibacterial agents of similar spectrum to (1)synergistically enhance treatment of severe Gram-negative infectionscovered by the spectrum of this compound or (2) add coverage in severeinfections in which multiple organisms are suspected in which anotheragent of a different spectrum may be required in addition to thiscompound. Potential agents include members of the aminoglycosides,penicillins, cephalosporins, fluoroquinolones, macrolides,glycopeptides, lipopeptides and oxazolidinones. The treatment caninvolve administering a composition having both active agents oradministration of the inventive compounds followed by or preceded byadministration of an additional active antibacterial agent.

Characterization and Purification Methods

Referring to the examples that follow, compounds of the presentinvention were characterized by high performance liquid chromatography(HPLC) using a Waters Millenium chromatography system with a 2690Separation Module (Milford, Mass.). The analytical columns were AlltimaC-18 reversed phase, 4.6×250 mm from Alltech (Deerfield, Ill.). Agradient elution was used, typically starting with 5% acetonitrile/95%water and progressing to 100% acetonitrile over a period of 40 minutes.All solvents contained 0.1% trifluoroacetic acid (TFA). Compounds weredetected by ultraviolet light (UV) absorption at either 220 or 254 nm.HPLC solvents were from Burdick and Jackson (Muskegan, Mich.), or FisherScientific (Pittsburgh, Pa.). In some instances, purity was assessed bythin layer chromatography (TLC) using glass or plastic backed silica gelplates, such as, for example, Baker-Flex Silica Gel 1 B2-F flexiblesheets. TLC results were readily detected visually under ultravioletlight, or by employing well known iodine vapor and other variousstaining techniques.

Mass spectrometric analysis was performed on one of two LCMSinstruments: a Waters System. (Alliance HT HPLC and a Micromass ZQ massspectrometer; Column: Eclipse XDB-C-18, 2.1×50 mm; solvent system: 5-95%(or 35-95%, or 65-95% or 95-95%) acetonitrile in water with 0.05% TFA;flow rate 0.8 mL/min; molecular weight range 500-1500; cone Voltage 20V; column temperature 40° C.) or a Hewlett Packard System (Series 1100HPLC; Column: Eclipse XDB-C18, 2.1×50 mm; solvent system: 1-95%acetonitrile in water with 0.05% TFA; flow rate 0.4 mL/min; molecularweight range 150-850; cone Voltage 50 V; column temperature 30° C.). Allmasses are reported as those of the protonated parent ions.

GCMS analysis was performed on a Hewlet Packard instrument (HP6890Series gas chromatograph with a Mass Selective Detector 5973; injectorvolume: 1 μL; initial column temperature: 50° C.; final columntemperature: 250 C; ramp time: 20 minutes; gas flow rate: 1 mL/min;column: 5% phenyl methyl siloxane, Model #HP 190915-443, dimensions:30.0 m×25 m×0.25 m).

Nuclear magnetic resonance (NMR) analysis was performed with a Varian300 Mhz NMR (Palo Alto, Calif.). The spectral reference was either TMSor the known chemical shift of the solvent. Some compound samples wererun at elevated temperatures (e.g. 75° C.) to promote increased samplesolubility.

The purity of some of the invention compounds was assessed by elementalanalysis (Desert Analytics, Tucson, Ariz.)

Melting points were determined on a Laboratory Devices Mel-Tempapparatus (Holliston, Mass.).

Preparative separations were carried out using a Flash 40 chromatographysystem and KP-Sil, 60A (Biotage, Charlottesville, Va.), or by flashcolumn chromatography using silica gel (230-400 mesh) packing material,or by HPLC using a C-18 reversed phase column. Typical solvents employedfor the Flash 40 Biotage system and flash column chromatography weredichloromethane, methanol, ethyl acetate, hexane, acetone, aqueoushydroxyamine and triethyl amine. Typical solvents employed for thereverse phase HPLC were varying concentrations of acetonitrile and waterwith 0.1% trifluoroacetic acid.

Compounds of the present invention can be readily synthesized using themethods described herein, or other methods, that are well known in theart. For example, the synthesis of hxdroxamic acids or similar scaffoldshaving a wide variety of substituents are comprehensively reviewed inKline, T., et al., “Potent, novel in vitro inhibitors of the Pseudomonasaeruginosa deacetylase LpxC” J. Med Chem. 2002, 45(14), 3112-29; U.S.Pat. No. 5,925,659; Pirrung, M. C., et al., “A Convenient Procedure forthe Preparation of Amino Acid Hydrokamates from Esters” J. Org. Chem.1995, 60, 8084-8085; Nhu, K., et al., “A New and Efficient Solid PhaseSynthesis of Hydroxamic Acids” J. Org. Chem. 1997, 62, 7088-7089;Internationa PCT Publication No. WO98/18754; Mellor, S. L., et al.,“N-Fmoc-aminoxy-2-chlortrityl Polystyrene Resin: A Facile Solid-phaseMethodology for the Synthesis of Hydroxamic Acids” Tetrahedron Lett.1997, 38, 3311-3314; Khan, S. I., et al., “A Facile and ConvenientSolid-phase Procedure for Synthesizing Nucleoside Hydroxamic Acids”Terahedron. Lett. 1998, 39, 8031-8034; Zhang, Y., et al., “Design,Combinatorial Chemical Synthesis, and in vitro Characterization of NovelUrea Based Gelatinase Inhibitors” Bioorg. Med. Chem. Lett. 1999, 9,2823-2826; Ito, Y., et al., “Synthetic Reactions by Complex Catalysts.XXXI, A Novel and Versatile Method of Heterocycle Synthesis” J. Am Chem.Soc. 1973, 95, 4447-4448; Ito, Y., et al., “Synthetic Reactions byComplex Catalysts XXXV” Syn. Commun. 1974, 4, 97-103; Witte, H., et al.,“Cyclische Imidsaurester aus Nitrilen and Aminoalkoholen” Liebigs Ann.Chem. 1974, 996-1009; Pattenden, G., et al., “Naturally Occurring LinearFused Thiazoline-Thiazole Containing Metabolites: Total Synthesis of (−)Didehydromirabazole A, a Cytotoxic Alkaloid from Blue-Green Algae” J.Chem. Soc. Perkin Trans 1993, 1, 1629-1636; Boyce, R. J., et al., “TotalSynthesis of Thiangazole, A Novel Naturally Occurring HIV-1 Inhibitorfrom Polyangium sp.” Tetrahedron 1995, 51, 7321-7330; Galeotti, N., etal., “Synthesis of Peptidyl Aldehydes from Thiazolines” Tetrahedron.Lett. 1997, 38, 2459-2462; Charette, A. B., et al., “Mild Method for theSynthesis of Thiazolines from Secondary and Tertiary Amides” J. Org.Chem. 1998, 63, 908-909; Bergeron, R. J., et al., “Effects of C-4Stereochemistry and C-4′ Hydroxylation on the Iron Clearing Efficiencyand Toxicity of Desferrithiocin Analogues” J. Med. Chem. 1999, 42,2432-2440; Raman, P., et al., “Titanium (IV)-mediated TandemDeprotection-cyclodehydration of Protected Cysteine N-Amides: BiomimeticSynthesis of Thiazoline- and Thiazole-containing Heterocycles” Org.Lett. 2000, 2, 3289-3292; Fernandez, X., et al., “Novel Synthesis of2-Thioazolines” Tetrahedron Lett. 2000, 41, 3381-3384; and Wipf, P., etal., “C. Thiolysis of Oxazolinenes: A New, Selective Method for theDirect Conversion of Peptide Oxazolines into Thiazolines” TetrahedronLett. 1995, 36, 6395-6398, which are incorporated herein by reference.

The synthesis of other non-hydroxamates compounds or more generally zincbinding groups are reviewed in Pirrung, M. C., et al., “Inhibition ofthe Antibacterial Target UDP-(3-O-acyl)-N-acetylglucosamine Deacetylase(LpxC): Isoxazoline Zinc Amidase Inhibitors Bearing Diverse MetalBinding Groups” J. Med. Chem. 2002, 45(19), 4359-4370; Jackman, J. E.,et al., “Antibacterial agents that target lipid A biosynthesis ingram-negative bacteria: inhibition of diverseUDP-3-O—(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylases bysubstrate analogs containing zinc binding motifs” J. Bio. Chem. 2000,275(15), 11002-11009; Brooks, C. D. W., et al, “Modulators ofLeukotriene Biosynthesis and Receptor Activation” J. Med. Chem. 1996,39(14), 2629-2654; Jeng, A. Y., et al., “Endothelin converting enzymeinhibitors” Current Pharmaceutical Design 1997, 3(6), 597-614; Zask, A.,et al., “Inhibition of matrix metalloproteinases: structure baseddesign” Current Pharmaceutical Design 1996, 2(6), 624-661; Skotnicki, J.S., et al., Current Opinion in Drug Discovery & Development 2003, 6(5),742-759.

The foregoing may be better understood by reference to the followingexamples, that are presented for illustration and not to limit the scopeof the inventive concepts.

EXAMPLES

The following are abbreviations used in the examples:

-   AcOH: Acetic acid-   aq: Aqueous-   ATP: Adenosine triphosphate-   Boc: tert-butoxycarbonyl-   Boc-Thr(OBn)-OH    3-(R)-Benzyloxy-2-(S)-tert-butoxycarbonylaminobutyric acid-   DAP or Dap: Diaminopropionate-   DCM:    4-(Dicyanomethylene)-2-methyl-6-(4-dimethylaminostyryl)-4H-pyran-   DEAD: Diethyl azodicarboxylate-   DIEA: Diisopropylethylamine-   DME: 1,2-dimethoxyethane-   DMF: N,N-Dimethylformamide-   DMSO: Dimethyl sulfoxide-   DPPA: Diphenyl phosphoryl azide-   Et₃N: Triethylamine-   EDC: N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide-   EDCI: 1-(3-dimethylaminopropyl)3-ethylcarbodiimide-   EtOAc: Ethyl acetate-   EtOH: Ethanol-   Fmoc: 9-fluorenylmethoxycarbonyl-   Gly-OH: glycine-   HATU: O-(7-azabenzotriaazol-1-yl)-N,N,N′N′=tetramethyluronium    hexafluorophophate-   HBTU: 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium    hexafluorophosphate-   Hex: hexane-   HOBt: butyl alcohol-   HOBT: 1-Hydroxybenzotriazole-   HPLC: High Pressure Liquid Chromatography-   IC₅₀ value: The concentration of an inhibitor that causes a 50%    reduction in a measured activity.-   iPrOH: Isopropanol-   LC/MS: Liquid Chromatography/Mass Spectrometry-   LRMS: Low Resolution Mass Spectrometry-   MeOH: Methanol-   NaOMe: sodium methoxide-   nm: Nanometer-   NMP: N-Methylpyrrolidone-   PPh₃: triphenyl phosphine-   RP-HPLC: Reversed-phase high-pressure liquid chromatography-   RT: Room temperature-   sat: Saturated-   TEA: Triethylamine-   TFA: Trifluoroacetic acid-   THF: Tetrahydrofuran-   Thr: Threonine-   TLC: Thin Layer Chromatography-   Trt-Br: Tert-butyl bromide

Nomenclature for the Example compounds was provided using ACD Nameversion 5.07 software (Nov. 14, 2001) available from Advanced ChemistryDevelopment, Inc. In addition, some of the compounds were named usingChemDraw Ultra 9.0 and ChemDraw Ultra 11.0 software available fromCambridgeSoft Corporation. Some of the compounds and starting materialswere named using standard IUPAC nomenclature.

Synthesis of N-Aryl Threonine Analogues and Formation of HydroxamateExample 1 Synthesis of3-bromo-4-fluoro-N-{(1S,2R)-2-hydroxy-1-[(hydroxyamino)carbonyl]propyl}benzamide(3) Preparation of(2S,3R)-2-(3-bromo-4-fluoro-benzoylamino)-3-hydroxy-butyric acid methylester (2)

Diisopropylethylamine (6.8 mL, 39.0 mmol) was added to a stirredsolution of 3-bromo-4-fluorobenzoic acid (1) (2.152 g, 9.83 mmol),L-threonine methyl ester hydrochloride (1.968 g, 11.6 mmol), EDCI (2.218g, 11.6 mmol) and HOBt (1.410 g, 10.4 mmol) in anhydrous DMF (60 mL) at0° C. under N₂. The solution was stirred at 0° C. for 1 h and at roomtemperature for 20 h. The solution was diluted with EtOAc (300 mL) andwashed with 1.0 M HCl (2×80 mL), saturated NaHCO₃ (2×80 mL), H₂O (4×80mL), dried over MgSO₄, filtered and concentrated in vacuo to give acolorless syrup which solidified on standing to afford 3.280 g (100%) of(2) as a white solid, mp 73-74° C. MS (ES+) m/z 333.9 (C₁₂H₁₃BrFNO₄+Hrequires 334.00).

Preparation of3-bromo-4-fluoro-N-{(1S,2R)-2-hydroxy-1-[(hydroxyamino)carbonyl]propyl}benzamide(3)

To a solution of hydroxylamine hydrochloride (66 mg, 0.95 mmol) inanhydrous MeOH (2.0 mL) at 0° C. under N₂ atmosphere was added sodiummethoxide (25 wt % in MeOH, 360 mg, 1.67 mmol). A precipitate formedimmediately and the cloudy white solution was stirred for 10 minutes at0° C. A solution of (2) (284 mg, 0.850 mmol) in MeOH (2.0 mL) was addedand the reaction stirred 2 h at 0° C. and then warmed gradually to roomtemperature overnight (17 h total). Aqueous 1.0 M HCl (10 mL) was addedand the solution extracted with 4:1 chloroform/isopropyl alcohol (4×20mL). The organic layers were combined, dried over Na₂SO₄ andconcentrated to give a pink foam. The crude solid was triturated withdiethyl ether (2×8 mL) and dried in vacuo to give (3) as a white foam:mp 152-153° C. Rf (10:1 CH₂Cl₂/MeOH on silica gel)=0.53.

Preparation of Hydroxamates Example 2 Synthesis of4-benzoyl-N-{(1S,2R)-2-hydroxy-1-[(hydroxyamino)carbonyl]propyl}benzamide(2)

To a solution of hydroxylamine hydrochloride (121 mg, 1.74 mmol) inanhydrous MeOH (2.0 mL) at 0° C. under N₂ atmosphere was added sodiummethoxide (25 wt % in MeOH, 680 mg, 3.14 mmol). A precipitate wasimmediately observed and the cloudy white solution was stirred for 10minutes at 0° C. A solution of methyl(2S,3R)-3-hydroxy-2-{[4-(phenylcarbonyl)phenyl]carbonylamino}butanoate(1) (534 mg, 1.56 mmol) in MeOH (3.0 nL) was added and the reactionstirred 3 h at 0° C., then warmed gradually to ambient temperatureovernight (18 h total). Aqueous 0.5 M HCl (20 mL) was added and thesolution extracted with 5:1 chloroform/isopropyl alcohol (4×40 mL). Theorganic layers were combined, dried over Na₂SO₄ and concentrated to givean orange foam. Purification by silica gel chromatography (increasingeluant polarity from 30:1 CH₂Cl₂/MeOH to 15:1 CH₂Cl₂/MeOH) afforded 228mg (43%) of (2).

Example 3 Synthesis of(2R,3R)-3-hydroxy-1-{[4-(trifluoromethoxy)phenyl]carbonyl}pyrrolidine-2-carbohydroxamicacid (3) Preparation of((2R,3R)-3-hydroxy-1-{[4-(trifluoromethoxy)phenyl]carbonyl)}pyrrolidin-2-yl)-N-(phenylmethoxy)carboxamide (2)

To a solution of (2R,3R)-3-hydroxy-1-{[4-(trifluoromethoxy)phenyl]carbonyl}pyrrolidine-2-carboxylic acid (1) (405 mg, 1.27 mmol),benzylhydroxylamine hydrochloride (243 mg, 1.52 mmol), HATU (556 mg,1.46 mmol), and HOBt (178 mg, 1.32 mmol) in DMF (10 mL) at 0° C. wasadded diisopropylethylamine (710 μL, 4.07 mmol) with stirring. Thecooling bath was removed after one hour and the reaction mixture stirredat ambient temperature for 18 h and then diluted with EtOAc (200 mL).The organic layer was washed with 1.0 M HCl (2×60 mL), sat. NaHCO₃ (2×60mL) and H₂O (5×60 mL), dried over MgSO₄ and concentrated to give 493 mg(92%) of (2), a colorless oil that slowly crystallized upon standing. Rf(25:1 CH₂Cl₂/MeOH)=0.35.

Preparation of(2R,3R)-3-hydroxy-1-{[4-(trifluoromethoxy)phenyl]carbonyl}pyrrolidine-2-carbohydroxamicacid (3)

To a solution of (2) (143 mg, 0.337 mmol) in EtOH (10 mL) was added 20%Pd(OH)₂/C (50 mg). The solution was purged with hydrogen gas (approx.0.5 L from a 1 L balloon) and then stirred under an atmosphere of H₂(balloon pressure). TLC analysis showed no starting material after onehour. The solution was diluted with EtOAc (10 mL) and filtered throughcelite, washing with 20:1 EtOAc/EtOH (50 mL) The solution wasconcentrated and dried in vacuo to afford 90 mg (80%) of (3) as a stickywhite foam: mp 64-65° C. Rf (10:1 CH₂Cl₂/MeOH)=0.29.

Synthesis of N-Benzyl Threonine Analogues by Reductive Amination Example4 Synthesis of(2S,3R-3-hydroxy-2-{[(4-phenylphenyl)methyl]amino}butanehydroxamic acid(3)

Triethyl amine (1.70 mL, 12.1 mmol) was added to a stirred suspension ofL-threonine methyl ester hydrochloride (1.030 g, 6.07 mmol) and4-biphenylcarboxaldehyde (1) (1.104 g, 6.06 mmol) in THF (25 mL). After20 min, NaBH(OAc)₃ (1.800 g, 8.49 mmol) was added and the suspensionstirred for 20 h. The reaction was monitored by TLC (50:1 DCM/MeOH,R_(f)=0.4). The reaction mixture was quenched with saturated NaHCO₃ (50mL), extracted with EtOAc (2×120 mL), dried over MgSO₄, filtered andconcentrated to give a yellow oil. Purification by silica gelchromatography (150:1 DCM/MeOH) afforded 1.220 g (67% yield, 98% pure)of (2) as a pale yellow oil. HPLC (260 nm, 34 min run) 14.2 min; LRMS(ES+) m/z 299.9 (C₁₈H₂1NO₃+H requires 300.10).

Compound (3) was then formed by the addition of NH₂OH in MeOH/NaOMe at0° C., warming to ambient temperature of the period of several hours.LCMS MH+301.15.

General Methods for Making Phenyl-Benzoic Acids and Phenyl-BenzoateEsters (See Example 5 Below)

Suzuki Procedures Using Pd(Dppf)Cl₂-DCM Catalyst and a THF/H₂O Mixture

Reagent MW Eq. g/ml mmol BromoArene #1 ~300 1 100 mg ~0.33 Boronic Acid#2 — 1.2 — ~0.40 Na₂CO₃ 105.99 3 104 m ~0.99 Pd(dppf)Cl₂ 816.63 0.1-0.227-54 mg ~0.033-0.066 THF (3) (sparged with 0.75 ml argon for 5 min.)water (1) (sparged with 0.25 ml argon for 5 min.)

1 eq aryl halide (1) was added to 1.2 eq. (2) and Pd(dppf)Cl₂ in THF,followed by addition of water and stirred 8 hours at RT. Upon completion(usually over night), the reactions are diluted with ethyl acetate (5-10ml) and water (1 ml). The organic layer is separated and washed withNaHCO₃ (2×3 ml), water (1×3 ml), brine (1×3 ml), dried with Na₂SO₄,filtered and concentrated in an 8 ml glass vial. The residue isdissolved in DMSO and injected on a preparatory HPLC reverse phasecolumn to afford >80% yield.

Suzuki Procedures Using Pd(Dppf)Cl₂-DCM Catalyst and DMF Solvent

Reagent MW Eq. g/ml mmol BromoArene #1 ~500 1 20 mg ~0.04 Boronic Acid#2 ~200 2 ~14 mg ~0.08 Pd(dppf)Cl₂ 816.63 0.25 10 mg ~0.01-0.02 TEA101.19 5 28 μL ~0.2  DMF (dry & sparged with 0.5 ml argon for 5 min.)

The haloarene (1) and boronic acid (2) were weighed out and placed inthe reaction flask. The DMF was sparged with argon for 5-10 minutes,followed by TEA addition, and the reaction was lightly bubbled withargon. The solid Pd(dppf)Cl₂ catalyst was added in one portion. The vialwas flushed with argon, capped tight and stirred or shaken at ˜80° C.Upon reaching completion (over night), the reaction was filtered andinjected on a preparatory HPLC reverse phase column (80% yield).

Synthesis of Methyl DAP Analogues Example 53-(R)-Amino-2-(S)-[(4′-ethyl-biphenyl-4-carbonyl)-amino]-butyl-hydroxamicacid (8) Preparation of N-triphenylmethyl allo-threonine Methyl Ester(2)

-   *For similar-procedures see: Righi, P., et al., B. Organic Letters    2002, 4(4), 497-500.

A solution of trityl bromide (3.2 g, 10.0 mmol) in CHCl₃ (40 ml) wasadded dropwise to a stirred solution of allo-threonine methyl ester HClsalt (1) (2.0 g, 12.0 mmol) and DIEA (5.2 ml, 30.0 mmol) in CHCl₃ (60ml) at rt under N₂. The reaction could be followed by TLC eluting withEtOAc/Hex (40:60) (Rf=0.3). After stirring 12 h, the reaction wasconcentrated to a brown oil. The crude product was diluted with EtOAc(170 ml) and washed with 0.2 N citric acid (2×50 ml), water (2×50 ml),brine (50 ml), dried (Na₂SO₄), filtered and concentrated under reducedpressure to yield 3.73 g (85% yield, 95% pure) of (2) as a yellow solid.HPLC (220 nm, 41 min. run) 30.90 min.; HPLC (220 nm, 17 min. run) 14.86min.; LCMS:LC (214 nm) 3.06 min., MS (ES+) m/z 376.2 (C₂₄H₂₅NO₃+Hrequires 376.18).

Preparation of 3-(R)-Azido-2-(S)-(trityl-amino)-butyric Acid MethylEster (3)

-   *For similar procedures see: Matsuda, A., et al., J. Med. Chem.    1991, 34, 999-1002.

A solution of pure DEAD (2.9 ml, 17.8 mmol) in THF (5 ml) was addedslowly dropwise to a stirred solution of trt-allo-threonine methyl ester(2) (4.1 g, 10.9 mmol) and PPh₃ (2.9 g, 10.9 mmol) in THF (40 ml) at 0°C. under N₂. After 3 min., a solution of DPPA (6.4 ml, 29.7 mmol) in THF(5 ml) was added to the orange-yellow reaction solution at 0° C. After 1h, the reaction was allowed to warm to rt. After 40 h, the reaction hadreached completion by TLC (Hexane/DCM/EtOAc (64:20:16) (Rf=0.6)) andLCMS. The yellow solution was concentrated to give 18 g of crudematerial that was purified by column chromatography eluting withHexane/EtOAc (88:12) giving 3.5 g of 70% pure product after evaporation.The product was purified again (to remove trityl alcohol and a crotylside-product formed during the reaction by elimination) by columnchromatography eluting with Hexane/DCM/EtOAc (76:20:4) giving 1.65 g(38% yield) of (3) as a pale yellow oil after concentration and dryingin vacuo. Note that the trityl protecting group would hydrolyze whenexposed to TFA while running the sample on HPLC.

Alternately, the reaction could be carried out in dry DCM. A reactionusing 5.44 g (14.5 mmol) of trt-allo-threonine methyl ester (2) in DCM(100 ml) with PPh₃ (3.8 g, 14.5 mmol), pure DEAD (3.4 ml, 21.8 mmol) inDCM (5 ml) and DPPA (6.3 ml, 24.0 mmol) in DCM (10 ml) were combinedfollowing the procedure above. After 3 days, the reaction did notprogress further by TLC and LCMS. After the same work up, 2.97 g of theproduct was obtained in 51% yield.

HPLC (220 nm, 41 min. run) 40.5 min.; HPLC (220 nm, 17 min. run) 16.32min.; LCMS: LC (214 nm) 3.7 min., MS (ES+) m/z 401.2 (C₂₄H₂₅N₃O₂+Hrequires 401.15).

Preparation of 2-(S)-Amino-3-(R)-azido-butyric acid methyl ester HClsalt (4)

A solution of Trt-Azido-Thr-OMe (3) (4.8 g, 12.0 mmol) was dissolved ina 95% TFA/DCM solution (60 ml) at rt with stirring. After 2.5 h, thereaction was complete by LCMS. The bright yellow solution was dilutedwith 0.5 N aq. HCl (300 ml). The aqueous layer was extracted with DCM(2×30 ml) and then lyophilized to dryness. The white solid was dissolvedin AcCN/water (50:50) (100 ml) and again lyophilized to dryness toproduce a consistent powder and remove as much of the TFA as possible.The azido-Thr product (4), 2.26 g (97% yield, 95% pure) of a whitesolid, was obtained as the HCl salt. HPLC (220 nm, 41 min. run) 7.91min.; HPLC (220 nm, 17 min. run) 3.36 min; LCMS: LC (214 nm) 0.48 min.,MS (ES+) m/z 159.3 (C₅H₁₀N₄O₂+H requires 159.08).

Preparation of3-(R)-Azido-2-(S-[(4′-ethyl-biphenyl-4-carbonyl)-amino]-butyric acidmethyl ester (6)

An EDC.HCl (249 mg, 1.3 mmol) was added to a stirred colorless solutionof azido-Thr-OMe.HCl (4) (195 mg, 1.0 mmol), HOBT (158 mg, 1.0 mmol),4′-Ethyl-biphenyl-4-carboxylic acid (5) (226 mg, 1.0 mmol) and DIEA(0.44 ml, 2.5 mmol) in DCM (10 ml) at rt under N₂. After 24 h, thereaction had reached completion by TLC (Hexane/EtOAc (60:40) (Rf=0.3))and LCMS. The reaction was evaporated under reduced pressure to a browntar. The crude product was dissolved in EtOAc (100 ml) and washed with0.2N aq. HCl (2×50 ml), aq. sat. NaHCO₃ (50 ml), brine (50 ml), dried(Na₂SO₄), filtered and concentrated under reduced pressure to yield acrude brown solid. The crude material was further purified by columnchromatography eluting with Hexane/EtOAc (70:30) giving 245 mg (67%yield) of pure product (6) after evaporation and drying in vacuo. HPLC(220 nn, 41 min. run) 33.87 min.; HPLC (220 nm, 17 min. run) 15.61 min;LCMS: LC (214 nm) 3.25 min., MS (ES+) m/z 367.2 (C₂₀H₂₂N₄O₃+H requires367.17).

Preparation of3-(R)-Amino-2-(S)-[(4′-ethyl-biphenyl-4-carbonyl)-amino]-butyric acidmethyl ester (7)

A solution of biphenyl azido-Thr methyl ester (6) (244 mg, 0.67 mmol) inMeOH (10 ml) was made by sonicating until the milky precipitate cleared.After bubbling nitrogen through the reaction solution for 30 sec., 10%Pd/C was added in one portion. The reaction was stirred under nitrogenat RT. The reaction was exposed to aspirator vacuum to remove thenitrogen and then opened to the hydrogen gas at balloon pressure (˜1atm). The reaction stirred for 3 h at which time the hydrogen wasexchanged for nitrogen. The reaction was filtered through a pad ofcelite to remove the palladium. The celite pad was washed with MeOH (30ml). The combined fractions of MeOH were evaporated under reducedpressure and dried in vacuo to give 225 mg (99% yield) of pure product(7) as a white solid. HPLC (220 nm, 17 min. run) 10.79 min.; LCMS: LC(214 nm) 2.21 min., MS (ES+) m/z 341.2 (C₂₀H₂₄N₂O₂+H requires 341.18).

Preparation of3-(R)-Amino-2-(S)-[(4′-ethyl-biphenyl-4-carbonyl)-amino]-butyl-hydroxamicacid (8)

To a stirred suspension of biphenyl-amino-Thr methyl ester (7) (225 mg,0.6 mmol) and hydroxylamine HCl salt (460 mg, 6.6 mmol) in MeOH (7 ml)and DCM (5 ml) was added fresh solid NaOMe powder (430 mg, 7.92 mmol) inone portion. After stirring for 2 min. at rt under nitrogen, the pH ofthe reaction on wet pH paper was approximately 7-8. The suspension hadchange from larger particles of white solid to a finely-divided milkyconsistency. The pH of the reaction was checked after adding smallportions of NaOMe (50-100 mg) and allowing 2 min. for the reaction toequilibrate. The pH of the reaction reached a stable 11-12 after thefinal portion of NaOMe was added (250 mg total). The reaction wasinitiated at pH 11 and proceeded quickly. After 30 min., the reactionreached 85% completion as determined by LCMS, and the reaction wasplaced in a −10° C. bath. The cold mixture filtered over fine filterpaper on a Buchner funnel. The white residue was washed with MeOH (15ml). The organic fractions were collected and concentrated under reducedpressure to give crude product (750 mg). The crude product (only one 150mg portion) was dissolved in DMSO (1 ml), AcCN (1000 and water (100 μl),passed through a Teflon syringe filter, and the clear filtrate wasinjected on a preparative HPLC. The purification used a 20×50 mm Ultro120 C18 column running a 22 ml/min 2% gradient (AcCN/water, 0.1% TFA)for 16 min. The purified fractions were lyophilized to dryness. Theproduct as the TFA salt was dissolved in AcCN/water (50:50) (5 ml), 1Naq. HCl (1 equivalent) and lyophilized again to give 11.5 mg of (8) as awhite powder as an HCl salt (23% yield). HPLC (220 nm, 41 min. run)19.31 min.; HPLC (220 nm, 17 min. run) 9.39 min; LCMS: LC (214 nm) 1.98min., MS (ES+) m/z 342.2 (C₁₉H₂₃N₃O₃+H requires 342.17).

Synthesis of 4′-Benzamide Biphenyl Threonine Hydroxamic Acid Example 6Biphenyl-4,4′-dicarboxylic acid 4′-[(3-Boc-amino-propyl)-amide]4-[((2R)-hydroxy-(1S)-hydroxycarbamoyl-propyl)-amide] (6) and Example 7Biphenyl-4,4′-dicarboxylic acid 4′-[(3-amino-propyl)-amide]4-[((2R)-hydroxy-(1S)-hydroxycarbamoyl-propyl)-amide] (7) Synthesis of(2S,3R)-2-amino-3-(phenylmethoxys)-N-(phenylmethoxy)butanamide (1)

To a suspension of benzylhydroxylamine hydrochloride (8.310 g, 52.06mmol), Boc-Thr(OBn)-OH (14.01 g, 45.28 mmol), EDCI (10.01 g, 52.21mmol), and HOBt (6.90 g, 51.06 mmol) in CH₂Cl₂ (300 mL) at 0° C. wasadded diisopropylethylamine (28.3 mL, 162 mmol) with stirring. Thecooling bath was removed after one hour and the reaction mixture stirredat ambient temperature for 20 h and was then diluted with CH₂Cl₂ (300mL). The organic layer was washed with 1.0 M HCl (2×200 mL), sat. NaHCO₃(2×200 mL) and brine (200 mL), dried over MgSO₄ and concentrated to give14.5 g of a white solid. The crude solid was treated with a solution oftrifluoroacetic acid (90 mL) in CH₂Cl₂ (90 mL) and stirred for 2.5 h.The reaction mixture was concentrated by rotary evaporation and thendiluted with CH₂Cl₂ (600 mL). The organic layer was washed with sat.NaHCO₃ (2×200 mL), dried over MgSO₄ and concentrated to give a darkorange oil. Purification by silica gel chromatography (50:1 CH₂Cl₂/MeOH)afforded (1) (8.9 g) as a pale yellow oil. Rf (50:1 CH₂Cl₂/MeOH onsilica gel)=0.2.

Preparation of(1S,2R)-4′-(2-benzyloxy-1-benzyloxycarbamoyl-propylcarbamoyl)-biphenyl-4-carboxylicacid (3)

To a suspension of 4,4′-biphenyldicarboxylic acid (2) (1.360 g, 5.61mmol) in DMF (180 mL) was added BOP (2.007 g, 4.54 mmol) and DIEA (1.7mL, 9.8 mmol). A solution of (1) (944 mg, 3.00 mmol) in DMF (20 mL) wasadded and the reaction stirred for 18 h. The solution was diluted withEtOAc (250 mL) and washed with 1.0 M HCl (500 mL). The aqueous layer wasextracted with EtOAc (250 mL) and the organic layers combined. Theorganic layer was washed with 1.0 M HCl (250 mL), dried over MgSO₄, andconcentrated to give a crude yellow solid. Purification by silica gelchromatography (60:1 CH₂Cl₂/MeOH) gave 210 mg (3) (13% yield) as ayellow solid. R_(f)=0.80 (10:1 CH₂Cl₂/MeOH); LRMS (ES+) m/z 539.1(C₃₂H₃₀N₂O₆+H requires 539.22).

Preparation of biphenyl-4,4′-dicarboxylic acid4′-[(3-(Boc)-amino-propyl)-amide]4-[(2R)-benzyloxy-(1S)-benzyloxycarbamoyl-propyl)-amide](5)

To a solution of (3) (200 mg, 0.371 mmol), EDCI (78 mg, 0.407 mmol), andHOBt (52 mg, 0.385 mmol) in DMF (2 mL) was added t-ButylN-(3-aminopropyl)carbamate (4) (71 mg, 0.407 mmol) and DIEA (180 μL, 1.0mmol). The reaction mixture was stirred 24 h, diluted with EtOAc (150mL), washed with 1.0 M HCl (2×60 mL), saturated NaHCO₃ (2×60 mL), H₂O(3×60 mL), dried over MgSO₄ and concentrated to give a crude whitesolid. Purification by silica gel chromatography (25:1 CH₂Cl₂/MeOH)afforded 194 mg (75% yield) of (5) as a white solid. R_(f)=0.15 (50:1CH₂Cl₂/MeOH); LRMS (ES+) m/z 695.2 (C₄₀H₄₆N₄O₇+H requires 695.35).

Preparation of Biphenyl-4,4′-dicarboxylic acid4′-[(3-Boc-amino-propyl)-amide]4-[((2R)-hydroxy-(1S)-hydroxycarbamoyl-propyl)-amide](6)

A solution of (5) (190 mg, 0.273 mmol) in THF (5 mL) and MeOH (3 mL) wascharged with Pd(OH)₂ (20%/C, 20 mg, 0.04 mmol) and stirred under ahydrogen atmosphere (balloon pressure) for 16 h. The crude mixture wasfiltered through a plug of celite eluting with 2:1 MeOH/THF (15 mL) andconcentrated to give an orange syrup. Purification by silica gelchromatography (5:1:1 THF/MeOH/CH₂Cl₂) afforded 110 mg (78% yield) of(6) as a white foam, mp 75-77° C. R_(f)=0.20 (10:1 CH₂Cl₂/MeOH); LRMS(ES+) m/z 515.4 (C₂₆H₃₄N₄O₇+H requires 515.26).

Preparation of Biphenyl-4,4′-dicarboxylic acid4′-[(3-amino-propyl)-amide]4-[((2R)-hydroxy-(1S)-hydroxycarbamoyl-propyl)-amide](7)

A flask containing (6) (80 mg, 0.155 mmol) was treated with 50%TFA/CH₂Cl₂ (6.0 mL) and stirred for 2.5 h. The reaction mixture wasconcentrated by rotary evaporation to give a brown syrup. Purificationby RP-HPLC (C₁₈ column, CH₃CN gradient 5-70%, 0.1% TFA, UV analysis 300nm, 36 min) and lyophilization of the collected fractions afforded 14 mg(21% yield) of (7) as a white solid. LRMS (ES+) m/z 415.3 (C₂₁H₂₆N₄O₅+Hrequires 415.20); RP-HPLC (300 nm, 36 min run) 18.2 min.

Example 8 Synthesis ofN-(2-(N-hydroxycarbamoyl)(2S)-2-{[4-(4-ethylphenyl)phenyl]carbonylamino}ethyl)acetamide(4) Preparation of3-Acetylamino-2-(9H-fluoren-9-ylmethoxycarbonylamino)-propionic acid (2)

Acetic anhydride (425 uL) in THF (5 ml) was added to a cloudy mixture ofFmoc-DAP-H (1) (980 mg, 3.0 mmol) and pyridine (483 uL, 6.0 mmol) in THF(15 ml) with stirring at rt. After 4 hours, the clear pale yellowsolution had reacted completely by LCMS. The reaction was evaporatedunder reduced pressure. The residue was dissolved in EtOAc (150 ml) andwashed with 0.1M NaHSO₄ (50 ml), water (50 ml), sat. brine (50 ml),dried with Na₂SO₄, filtered and concentrated under reduced pressure togive 1.1 g of crude product as a white solid. The crude product waspurified by prep. HPLC to give 0.99 g (90% yield) of (2).

Preparation of (2-Acetylamino-1-hydroxycarbamoyl-ethyl)-carbamic acid9H-fluoren-9-ylmethyl ester trityl resin (3)

A solution of (2) (980 mg, 0.56 mmol), HATU (0.146 g, 0.56 mmol) in NMP(1.7 ml) was made. After 2 min. of shaking, the activated acid was addedto the deprotected H₂N—O-Trt Resin (120 mg, 0.113 mmol) at rt withshaking. Deprotection of the Fmoc group from the resin was accomplishedusing 20% piperizine in DMF (4 ml) for 2 hours twice. The resin wasdrained and washed with DMF (2×5 ml) and DCM (2×5 ml).] After shakingfor 20 hours, the reaction was drained and washed with DMF (2×5 ml) andDCM (2×5 ml). The resin (3) was dried and used as is in the nextreaction.

Preparation ofN-(2-(N-hydroxycarbamoyl)(2S)-2-{[4-(4-ethylphenyl)phenyl]carbonylamino}ethyl)acetamide (4)

Resin (3) was treated with 20% piperizine in DMF (4 ml) for 2 hourstwice. The resin was drained and washed with DMF (2×5 ml) and DCM (2×5ml). The resin was dried in vacuo. A solution of4′-Ethyl-biphenyl-4-carboxylic acid (91 mg, 0.4 mmol), HATU (152 g, 0.4mmol) in NMP (1.0 ml) was made. After 2 min. of shaking, the activatedacid was added to the deprotected H-DAP(Ac)-Trt resin (120 mg, 0.113mmol) at rt with shaking. After shaking for 18 hours, the reaction wasdrained and washed with DMF (2×5 ml) and DCM (2×5 ml). The resin wasdried in vacuo. The product was cleaved from the resin through treatmentwith a solution of TFA (500 uL), DCM (500 uL) and water (50 uL) for 25min. The resin was filtered and washed with fresh DCM (2 ml). Thecombined TFA and DCM fractions are evaporated under reduced pressure.The residue was diluted with CH₃CN/water (1:1) (10 ml) and lyophilized.The crude product was purified by prep. HPLC. The crude product wasdissolved in DMSO (1 ml), passed through a Teflon syringe filter, andthe clear filtrate was injected on a preparative HPLC. The purificationused a 20×50 mm Ultro 120 C18 column running a 22 ml/min 2% gradient(AcCN/water, 0.1% TFA) for 16 min. The purified fractions werelyophilized to dryness. The solid residue was lyophilized again fromCH₃CN/water (1:1) (5 ml) give 8.6 mg of pure product (4) (˜21% yield).

Example 9 Synthesis of 4′-Ethyl-biphenyl-4-carboxylic acid(1-hydroxycarbamoyl-2-methanesulfonylamino-ethyl)-amide (3) Preparationof 4′-Ethyl-biphenyl-4-carboxylic acid(2-amino-1-hydroxycarbamoyl-ethyl)-amide trityl resin (2)

Pd(PPh₃)₄ (438 mg, 0.35 mmol) was added to a vial containingbiphenyl-DAP(Alloc)-Trt Resin (1) (500 mg, 0.35 mmol), dimethylbarbituric acid (600 mg, 3.5 mmol) and PPh₃ (438 mg, 0.35 mmol) in DCM(11 ml) at rt under argon. The mixture was sparged with argon and shakenfor 16 hours. The bright yellow mixture was drained and washed with DMF(8×10 ml) and DCM (8×10). The resin was dried in vacuo to give thedeprotected DAP resin (2).

Preparation of 4′-Ethyl-biphenyl-4-carboxylic acid(1-hydroxycarbamoyl-2-methane sulfonylamino-ethyl)-amide (3)

Methanesulfonyl chloride (85 uL, 1.1 mmol) was added to a mixture ofdeprotected DAP resin (2) (160 mg, 0.11 mmol) and lutidine (190 uL, 1.6mmol) in DCM (1.5 ml). After shaking for 16 hours, the mixture wasdrained and washed with DMF 10×2 ml) and DCM (5×2 ml). The product wascleaved from the resin through treatment with TFA/water (4:1) (1.5 ml).After shaking for 45 min., the TFA solution was collected from the resinby filtration, and the resin was washed with TFA (1 ml) and TFA/water(1:1) (10 ml). The combined TFA fractions were concentrated underreduced pressure to a reddish-brown solid. The product, identified byLCMS, was purified by prep. HPLC using a 20×50 mm Ultro 120 C18 columnrunning a 22 ml/min 4% gradient (AcCN/water, 0.1% TFA) for 16 min. Thepurified fractions were lyophilized to dryness. The solid residue waslyophilized again from CH₃CN/water (1:1) (5 ml) give 4 mg of pureproduct as a white solid (3) (˜9% yield).

Example 10 Synthesis of 4′-Ethyl-biphenyl-4-carboxylic acid[2-(3,3-dimethyl-ureido)-1-hydroxycarbamoyl-ethyl]-amide (3) (Continuedfrom (2) of Example 9 above)

Dimethylcarbamyl chloride (103 mg, 0.96 mmol) was added to a mixture ofdeprotected DAP resin (2) (125 mg, 0.096 mmol) and lutidine (225 uL,1.92 mmol) in DCM (1.5 ml). After shaking at rt for 5 hours, the mixturewas drained and washed with DCM (5×2 ml), DMF (5×2 ml) and DCM (5×2 ml).The product was cleaved from the resin through treatment with TFA/water(4:1) (1.5 ml). After shaking for 45 min., the TFA solution wascollected from the resin by filtration, and the resin was washed withTFA/water (1:1) (2 ml). The combined TFA fractions were concentratedunder reduced pressure to a reddish-brown solid. The product, identifiedby LCMS, was purified by prep. HPLC using a 20×50 mm Ultro 120 C18column running a 22 mvmin 4% gradient (AcCN/water, 0.1% TFA) for 16 min.The purified fractions were lyophilized to dryness. The solid residuewas lyophilized again from CH₃CN/water (1:1) (5 ml) give 5 mg of pureproduct as a white solid (3) (˜13% yield).

Example 11 Synthesis of 4′-Ethyl-biphenyl-4-carboxylic acid[2-(2-amino-ethylamino)-1-hydroxycarbamoyl-ethyl]-amide (2)

NaBH₃CN (3.1 mg, 0.05 mmol) followed by acetic acid (6 uL, 1.0 mmol)were sequentially added to a stirred suspension ofbiphenyl-DAP-hydroxamate (1) (20 mg, 0.096 mmol) andBoc-amino-acetaldehyde (6.4 mg, 0.4 mmol) in MeOH (1.5 ml) in a 4 mlvial. The reaction was followed by LCMS. After stirring 12 hours, thecloudy reaction was only 50% complete. The reaction was concentratedunder reduced pressure to a thick slurry that was dissolved in DMSO. Theproduct was purified by prep. HPLC using a 20×50 mm Ultro 120 C18 columnrunning a 22 ml/min 3% gradient (AcCN/water, 0.1% TFA) for 16 min. Thepurified fractions were lyophilized to dryness. The dried powder wasdissolved in CH₃CN/water (1:1) (1 ml) and 1M HCl (700 uL). After heatingat 50° C. for 75 min., the reaction mixture was again lyophilized todryness to produce 7.1 mg of (2) as a 2×HCl salt white powder (˜17%yield).

Example 12 Synthesis ofN-(1-(N-hydroxycarbamoyl)1S,2R)-2-hydroxypropyl)[4-(2-phenylethynyl)phenyl]carboxamidePreparation of 4-Phenylethynyl-benzoic acid (3)

4-iodo-benzoic acid methyl ester (1) (20.0 g, 76.34 mmol),ethynyl-benzene (2) (8.56 g, 83.96 mmol), PdCl₂(PPh₃)₂ (0.65 g, 0.92mmol), and CuI (0.35 g, 1.83 mmol) were mixed with THF (110 ml) in around bottom under argon. The dry THF was sparged with dry, oxygen-freeargon for at least 5 min. immediately before use. The reaction wascooled to 10° C. and TEA (16 ml) was added. The cooling bath was removedand the reaction was stirred at RT under argon. After 2.5 h, thereaction was diluted with EtOAc (400 ml) and the solids were filteredoff through a pad of celite. The organic filtrate was washed with 1M HCl(60 ml), sat. aq. NaHCO₃ (60 ml), water (60 ml), brine (60 ml), driedwith Na₂SO₄, filtered and concentrated under reduced pressure. The crudesolid methyl ester was dissolved in MeOH (400 ml), 6M NaOH (30 ml) andwater (50 ml). The reaction was stirred at 70° C. until a clear solutionwas formed (about 1 h). The reaction could be followed by LCMS. Thereaction was cooled and diluted with water (500 ml) and hexane (100 ml).The pH was adjusted to pH 6-7. The white solid that formed was collectedand washed with water (3×60 ml) and hexane (3×60 ml). The solid (3) wasdried in vacuo yielding 17.3 g (approximately quantitative yield in 99%purity).

Preparation of 3-hydroxy-2-(4-phenylethynyl-benzoylamino)-butyric acidmethyl ester (4)

A solution of threonine methyl ester (1.66 g, 9.8 mmol) and DIEA (1.53ml, 8.8 mmol) in DMF (10 ml) was added to a stirred solution of (3)(1.55 g, 7.0 mmol) and DIEA (1.53 ml, 8.8 mmol) in DMF (11 ml) at rt.After 12 h, the reaction was diluted with EtOAc (300 ml) and washed with0.5M HCl (2×60 ml), sat. aq. NaHCO₃ (60 ml), 50% diluted brine (60 ml),sat. brine (60 ml), dried with Na₂SO₄, filtered and concentrated underreduced pressure. Upon drying in vacuo, 2.34 g of white solid (4) wasobtained (approximately quantitative yield in 99% purity).

Preparation ofN-(2-Hydroxy-1hydroxycarbamoyl-propyl)-4-phenylethynyl-benzamide (5)

A solution of (4) (2.34 g, 7.0 mmol) in MeOH (20 ml) and DCM (30 ml) wasadded to a cooled (−10° C. bath) suspension of hydroxylamine HCl salt(4.81 g, 70.0 mmol) and NaOMe (4.16 g, 77.0 mmol) in MeOH (30 ml).Follow reaction by LCMS. After stirring for 2 hours, the reaction seemsto stall at 50% completion. Add an additional 1 equivalent of NaOMe(0.416 g). After 3 hours, the reaction was 75% complete. Add anadditional 0.5 equivalent of NaOMe (0.21 g). After 4 hours, the reactionwas 90% complete. Add an additional 0.15 equivalent of NaOMe (0.064 g)for a total of 12.65 equivalents of NaOMe. The pH of the reaction wasbetween 11-12 and had reacted about 95% completion. The reaction wasdiluted with EtOAc (500 ml) and washed with sat. aq. NaHCO₃ (2×60 ml),50% diluted brine (60 ml), sat. brine (60 ml), dried with Na₂SO₄,filtered and concentrated under reduced pressure. The residue wasdissolved in minimal DMA. The product was purified by prep. HPLC using areverse phase Ultro 120 C18 column running a 2% gradient (AcCN/water,0.1% TFA). The purified fractions were lyophilized to dryness. Theproduct as the TFA salt dissolved in AcCN/water (50:50) (80 ml), 1N aq.HCl (13 equivalent) and lyophilized again to give 1.3 g of white powder(5) in 55% yield and >97% purity.

Example 13 Synthesis of3-(R)-Amino-2-(S)-(3-phenylethynyl-benzoylamino)-butyl-hydroxamic acid(10) Preparation of3-(R)-Azido-2-(S)-(3-phenylethynyl-benzoylamino)-butyric acid methylester (9)

Compound (9) was made by the same procedures as for compound (6) inExample 5 above using compound (3) from Example 12 above. The product(9) was obtained in 92% yield (952 mg). HPLC (220 nm, 41 min. run) 32.64min.; HPLC (220 nm, 17 min. run) 15.08 min LCMS: LC (214 nm) 3.16 min.,MS (ES+) m/z 363.1 (C₂₀H₁₈N₄O₃+H requires 363.14).

Preparation of3-(R)-Amino-2-(S)-(3-phenylethynyl-benzoylamino)-butyl-hydroxamic acid(10)

Triphenylphosphine (526 mg, 2.0 mmol) was added to a stirred solution of(9) (726 mg, 2.0 mmol) at rt. After 3 days the reaction reachedcompletion as judged by TLC (EtOAc/Hex (2:1)) and LCMS. The reaction wasconcentrated under reduced pressure to give an ivory colored solid. Thecrude amino-phosphine was dissolved in MeOH (20 ml) to give a paleyellow solution. To the solution of amino-phosphine was addedsequentially hydroxylamine HCl salt (1.4 g, 20.0 mmol) followed by freshsolid NaOMe powder (1.3 g, 24.0 mmol) to make a milky pH 10 suspension.After 36 h, the reaction was complete by LCMS. The reaction wasevaporated under reduced pressure to give a yellow solid that was driedin vacuo. The crude product (2.75 g) was triturated with ether (3×50 ml)to remove impurities (P(O)Ph₃) and then was dissolved in abs. EtOH (120ml) with sonication for 15 min. A fine white powder was suction filteredoff, and the clear yellow ethanolic portion was concentrated to a smallvolume. The crude product was dissolved in DMSO (8 ml) and purified bypreparative HPLC (Ultro 120 C18 75×300 mm column) running a gradient(AcCN/water, 0.1% TFA) from 5 to 70% for 55 min. The purified fractionswere pooled together and lyophilized to dryness. The product as the TFAsalt was dissolved in AcCN/water (50:50) (100 ml), IN aq. HCl (1equivalent) and lyophilized again to give 325 mg of (10) as a lightyellow powder as the HCl salt (43% yield). HPLC (220 nm, 41 min. run)18.31 min.; HPLC (220 nm, 17 min. run) 9.11 min; LCMS: LC (214 nm) 1.91min., MS (ES+) m/z 338.1 (C₁₉H₁₉N₃O₃+H requires 338.14).

Synthesis of 4′-(N-Acylamnino)-Tolan Dap Analogs Example 14 Synthesis of4-({4-[(aminoacetyl)amino]phenyl}ethynyl)-N-[(1S)-1-(aminomethyl)-2-(hydroxyamino)-2-oxoethyl]benzamidePreparation of 2-N-Boc-amino-N-(4-iodo-phenyl)-acetamide (2)

A solution of Boc-Gly-OH (1.752 g, 10.0 mmol) in DCM (18 mL) and DMF (1mL) was treated with EDCI (1.994 g, 10.4 mmol) and HOBt (1.351 g, 10.0mmol). After stirring 15 min, 4-iodoaniline (1) (2.290 g, 10.4 mmol) wasadded and the reaction monitored by TLC (25:1 DCM/MeOH (R_(f)=0.6)).After 24 h the solution was diluted with EtOAc (250 mL), washed with 1.0M HCl (3×100 mL), sat. NaHCO₃ (3×100 mL), brine (3×100 mL), dried overMgSO₄, filtered and concentrated in vacuo to afford 2.900 g (77% yield)of (2) as a white solid.

Preparation of (2S)-3-N-Boc-amino-(4-ethynyl-benzoylamino)-propionicacid methyl ester (4)

Diethylamine (3.5 mL, 20.0 mmol) was added to a stirred solution of4-ethynylbenzoic acid (3) (910 mg, 6.22 mmol), H-Dap(Boc)-OMehydrochloride (1.903 g, 7.47 mmol), EDCI (1.432 g, 7.47 mmol), and HOBt(910 mg, 6.73 mmol) in DMF (50.0 mL). After stirring 20 h, the reactionmixture was diluted with EtOAc (400 mL), washed with 1.0 M HCl (2×100mL), saturated NaHCO₃ (2×100 mL), H₂O (4×100 mL), dried over MgSO₄,filtered and concentrated in vacuo to give 2.140 g (99% yield) of (4) asa tan solid, mp=110-111° C. LRMS (ES+) m/z 346.9 (C₁₈H₂₂N₂O₅+H requires347.10).

Preparation of Methyl (2S)-3-[(tert-butoxy)carbonylamino]-2-({4-[2-(4-{2-[(tert-butoxy)carbonylamino]acetylamino}phenyl)ethynyl]phenyl}carbonylamino) propanoate (5)

To a suspension of methyl(2S)-3-[(tert-butoxy)carbonylamino]-2-[(4-ethynylphenyl)carbonylamino]propanoate(4) (200 mg, 0.577 mmol) and2-[(tert-butoxy)carbonylamino]-N-(4-iodophenyl)acetamide (2) (476 mg,1.26 mmol) was added Et₃N (350 μL, 2.5 mmol). The solution was purgedwith a stream of N₂ for several minutes and PdCl₂(PPh₃)2 (20 mg, 0.028mmol) and CuI (10.6 mg, 0.055 mmol) were added. The reaction mixture wasstirred at ambient temperature for 22 h and then concentrated by rotaryevaporation. The crude black residue was chromatographed twice by silicagel chromatography (30:1 CH₂Cl₂/MeOH) to give 285 mg (83%) of (5) as ayellow foam.

Preparation ofN-(4-{2-[4-(N-{1-(N-hydroxycarbamoyl)(1S)-2-[(tert-butoxy)carbonylamino]ethyl}carbamoyl)phenyl]ethynyl}phenyl)-2-[(tert-butoxy)carbonylamino]acetamide(6)

To a solution of hydroxylamine hydrochloride (98 mg, 1.41 mmol) in MeOH(1.3 mL) at 0° C. was added 25 wt % NaOMe (460 mg, 2.13 mmol). Thesolution was stirred at 0° C. for 15 min and then charged with asolution of (5) (279 mg, 0.469 mmol) in THF (1.5 mL) and MeOH (0.6 mL).The reaction was stirred at 0° C. for 30 min and at room temperature for2.5 h. The reaction mixture was diluted with 4:1 CHCl₃/iPrOH (50 ml) andwashed with 0.1 M HCl (30 mL). The layers were separated and the aqueouslayer extracted once more with 4:1 CHCl₃/iPrOH (30 ml). The organiclayers were combined, dried over Na₂SO₄, filtered and concentrated. Thecrude residue was suspended in 10:1 CH₂Cl₂/MeOH (4 mL), filtered, andwashed with 50:1 CH₂Cl₂/MeOH (2 mL) and Et₂O (10 mL) to afford 180 mg(64%) of (6) as a white powder.

Preparation of4-({4-[(aminoacetyl)amino]phenyl}ethynyl)-N-[(1S)-1-(aminomethyl)-2-(hydroxyamino)-2-oxoethyl]benzamide(7)

To an oven-dried flask containing (6) (130 mg, 0.218 mmol) was added 1:1TFA/CH₂Cl₂ (2.5 mL). The resulting pink solution was stirred for 2 h andconcentrated to give a pink gum. The crude residue was rinsed withCH₂Cl₂ (4 mL), concentrated by rotary evaporation and dissolved in THF(2 mL) and MeOH (0.4 mL). A solution of 4 M HCl in dioxane (200 μL) wasadded and the resulting precipitate filtered and washed with Et₂O (10mL) to afford 90 mg of (7) as a pale tan powder.

Reaction of Iodoaniline with Bromoacetyl Bromide

Bromoacetyl bromide (175 μL, 2.00 mmol) was added dropwise over 5minutes to a solution of 4-iodoaniline (438 mg, 2.00 mmol) and Et₃N (280μL, 2.00 mmol) in benzene (5 mL). The reaction was stirred 1 hour,treated with morpholine (1.0 mL, 11.5 mmol) and stirred overnight. Thereaction mixture was diluted with EtOAc (200 mL), washed with aqueous0.1 M KOH (50 nmL), H₂O (50 mL), dried over MgSO₄ and concentrated togive a yellow oil. Purification by silica gel chromatography (100:1CH₂Cl₂/MeOH) afforded 630 mg (91%) ofN-(4-iodophenyl)-2-morpholin4-ylacetamide as a waxy tan solid. Thisproduct was converted to analogues in a similar manner as Example 14.

Example A Preparation of4-[4-(6-Chloro-pyridin-3-yl)-buta-1,3-diynyl]-benzoic acid methyl ester

DIEA (9.7 ml, 55.1 mmol) was added to a stirred solution of4-iodo-benzoic acid (5.49 g, 22.2 mmol), HOAT (3.08 g, 22.6 mmol), EDC(4.33 g, 22.6 mmol) in DMF (85 ml). After 2 min., H-DAP(Boc)-OMe (1) wasadded in one portion. After 12 hours, the reaction was found complete byLCMS. The reaction was diluted with EtOAc/hexane (1:1) (500 ml). Theorganic phase-was washed with 1N HCl (2×80 ml), IN NaOH (2×80 ml), water(2×80 ml), sat. brine (80 ml), dried with Na₂SO₄, filtered andconcentrated under reduced pressure to give crude product. The residuewas filtered through a filter plug of silica eluting with EtOAc/hexane(1:1). The fractions with product were evaporated to give 9.3 g ofproduct (3-tert-Butoxycarbonylamino-2-(4-iodo-benzoylamino)-propionicacid methyl ester) in 93% yield. This product was converted to analoguesin a similar manner as the aforementioned Examples.

Example 15N-(1-(N-hydroxycarbamoyl)(1S,2R)-2-hydroxypropyl)(4-{2-[4-(morpholin4-ylmethyl)phenyl]ethynyl}phenyl)carboxamide(5) Preparation of (2S,3R)-2-[4-(4-formyl-phenylethynyl)-benzoylaminol-3-hydroxy-butyric acidmethyl ester (3)

A solution of (2S,3R)-methyl-2-(4-ethynylbenzamido)-3-hydroxybutanoate(1) (745 mg, 2.85 mmol), 4-iodobenzaldehyde (2) (902 mg, 3.89 mmol), andEt₃N (900 μL, 6.5 mmol) in THF (50 mL) was purged with a stream of N₂for two minutes and then treated with PdCl₂(PPh₃)₂ (70 mg, 0.10 mmol)and CuI (34 mg, 0.18 mmol). The reaction mixture was stirred 40 h,concentrated by rotary evaporation and purified by silica gelchromatography (40:1 DCM/MeOH) to give 0.833 g (80% yield) of (3) as apale yellow powder, mp=143-144° C. R_(f)=0.3 (25:1 DCM/MeOH); LRMS (ES+)m/z 366.1 (C₂₁H₁₉NO₅+H requires 366.13); HPLC (300 nm, 47 min) 15.3 min.

Preparation of (2S,3R)-3-Hydroxy-2-[4-(4-morpholin4-ylmethyl-phenylethynyl)-benzoylamino]-butyricacid methyl ester (4)

Sodium triacetoxyborohydride (0.670 g, 3.16 mmol) was added to asolution of benzaldehyde (3) (0.822 g, 2.25 mmol) and morpholine (260μL, 2.97 mmol) in THF (15 mL) under N₂ atmosphere and the reactionmonitored by TLC (25:1 DCM/MeOH, R_(f)=0.2). After stirring 4 h, thereaction mixture was quenched with saturated NaHCO₃ (150 mL), extractedwith EtOAc (3×100 mL), dried over MgSO₄, filtered and concentrated togive a yellow syrup. Purification by silica gel chromatography (35:1DCM/MeOH) afforded 0.844 g (86% yield) of (4) as a sticky white foam.

Preparation of (2S,3R)—N-(2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(5)

Sodium methoxide (25 wt % in MeOH, 1.860 g, 8.60 mmol) was added to astirred solution of hydroxylamine hydrochloride (400 mg, 5.76 mmol) inanhydrous MeOH (5 mL) at 0° C. under N₂ atmosphere. After stirring 20min, a solution of methyl ester (4) (829 mg, 1.90 mmol) in 1:1 MeOH/THF(6 mL) was added and the reaction mixture stirred at 0° C. for 1 h andat room temperature for 4 h. The reaction was quenched with 1.0 M HCl (6mL), concentrated by rotary evaporation to remove organic solvents, anddiluted with DMSO (4 mL). Analytical RP-HPLC (C₁₈ column, CH₃CN gradient5-35%, 0.1% TFA, UV analysis 300 nm, 16 min) indicated a purity of 85%for the crude product mixture. Purification by preparative RP-HPLC andlyophilization of the collected fractions gave 701 mg (81%) of (5) as afluffy white solid. LRMS (ES+) m/z 438.1 (C₂₄H₂₇N₃O₅+H requires 438.20);RP-HPLC (300 nm, 16 min run) 8.7 min.

Resin Procedures for Synthesizing Tolanyl Hydroxamates Example 16Synthesis of4-[(4-{[(benzylamino)acetyl]amino}phenyl)ethynyl]-N-{(1S,2R)-2-hydroxy-1-[(hydroxyamino)carbonylipropyl}benzamide

1. Coupling to Fmoc Hydroxylamine Resin

The resin was pre-swelled by adding DCM and shaking for 30 min. Theresin was drained, 20% piperdine was added in DMF, the resin was shaken1.25 hours, and finally drained and washed in 2×DMF and 2×DCM. Afterdraining completely, 20% piperdine in DMF was added to attain cleavagein 1.25 hours. The resin was washed 4×DMF, 4×DCM and drained completely.In a separate flask, the amino acid (Fmoc-Thr tBu-OH, or Fmoc-DAPBoc-OH, 4 eq) was mixed, HATU (4 eq), DMF (60 ml) and Hunig's (8 eq)base were added and stirred for 2-3 min. The mixture was added to theresin and shaken 20-24 hours. Subsequently, the resin was drained andrun with a standard wash (1×DCM, 4×DMF and 4×DCM). The Fmoc was removedfrom the amino acid by adding 20% piperdine in DMF and shaken 1.25hours, drained, and given the standard wash (1×DCM, 4×DMF and 4×DCM).

2. Coupling of 4-Iodobenzoic Acid to Amino Acid Resin

A mixture of 4-iodobenzoic acid (4 eq), HBTU (4 eq), DMF (60 ml) wasshaken for several minutes. Hunig's base (8 eq) was subsequently addedand the mixture was shaken further for 2-3 min. The pre-activatedmixture was then added to the prepared Thr or DAP resin (Fmoc removed,7.5 g, 5.775 mmol). The reaction is shaken 12-16 hours followed by thestandard wash (1×DCM, 4×DMF and 4×DCM).

3. Alkyne Coupling on Resin

To the 4-iodobenzoic resin (4 g, 3.08 mmol) was added4-aminophenylacetylene (3 eq), Pd(PPh₃)₂Cl₂ (0.04 eq), CuI (0.08 eq) andTHF (purged with Argon). After mixing for 1 min., TEA (4.5 eq) was addedand the reaction was shaken 12 hours at RT under argon.

4. Aniline Coupling with Bromoacetyl Chloride on Resin

To aniline resin (4 g, 3.08 mmol) was added DCM (30 ml) lutidine (10 eq)and shaken for 1 min. Bromoacetyl chloride (8 eq) in DCM (5 ml) wasadded slowly. After the addition, the slurry was shaken for 1.5 to 1.75hours. Subsequent draining and a wash with 2×DCM, 4×DMF and 4×DCM wasthen performed.

5. Displacement with Amines on Resin

To the bromoacetyl resin (125 mg), was added NMP (1.5 ml) followed byamine (0.2 g or ml, ie excess) and the slurry was shaken for 12-16 hoursat RT. To neutralize the salt, TEA was added. The imidazole was heatedat 38° C. for 24 h (in the case of anilines, they were heated at 38° C.for 48 h). The reaction mixture was drained and washed 4×DMF and 4×DCM.

6. Cleavage from Resin and Deprotection of Thr tBu and DAP Boc

The resin (125 mg) was soaked in TFA/water (80:20 v/v) (1.5 ml) at RTfor 45 min. Upon cleavage the solution was collected and the resin waswashed with more TFA/water mixture (0.75 ml). To the TFA/productsolution was added acetonitrile/water solution (1:1 v/v, 10 ml) and purewater (2.5 ml). The mixture was frozen in liquid nitrogen for 15 min andlyophilized. The dry residue was dissolved in the acetonitrile/watersolution (1:1 v/v, 10 ml) again followed by addition of 1M aq. HCl (1.2eq per basic nitrogen), frozen, and lyophilized to a powder.

Synthesis of 3′-Nitro-Tolan Threonine Hydroxamic Acid Example 17(1S,2R)—N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-4-(3-nitro-phenylethynyl)-benzamidePreparation of(1S,2R)—N-(2-tert-butoxy-1-hydroxycarbamoyl-propyl)-4-ethynyl-benzamideon Hydroxylamine 2-chlorotrityl Resin (3)

Fmoc-threonine resin (1) (0.522 g, 0.365 mmol, 0.70 mmol) was swelled inDCM (5 mL) for 2 h and drained. The resin was treated with 20%piperidine in DMF (6 mL) for 1 hour, washed with DMF (4×6 mL) and DCM(4×6 mL) and drained completely. In a separate flask, 4-ethynylbenzoicacid (2) (0.160 g, 1.10 mmol), DIC (0.280 mL, 1.79 mmol), HOBt (0.148 g,1.10 mmol) and DIEA (0.4 mL, 2.30 mmol) were dissolved in DCM (1 mL) andDMF (4 mL), stirred 15 min and added to the resin. After shaking for 36h, the mixture was drained, washed with DMF (4×6 mL) and DCM (4×6 mL)and dried in vacuo to give 0.495 g of (3) as a yellow resin.

Preparation of(1S,2R)—N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-4-(3-nitro-phenylethynyl)-benzamide(5)

Resin (3) (100 mg, 0.070 mmol) was swelled in DCM (2 mL) for 1 h anddrained. A solution of 1-iodo-3-nitrobenzene (4) (87.1 mg, 0.350 mmol)and Et₃N (150 μL, 1.10 mmol) in DMF (1.5 mL) was purged with a stream ofN₂ bubbles for two minutes and added to the resin. After mixing for 5min, PdCl₂(PPh₃)₂ (10.0 mg, 0.014 mmol) and CuI (7.0 mg, 0.036 mmol)were added and the mixture shaken for 26 h. The resin was drained,washed with DMF (3×2 mL), DCM (3×2 mL) and cleaved with 10% TFA/DCM (1.5mL) for 20 min. The solution was collected and the resin was rinsed withadditional 10% TFA/DCM (1.0 mL). The cleavage fractions were combined,treated with neat TFA (2.0 mL), stirred for 1 h at rt and concentratedby rotary evaporation to give a crude brown residue. Purification byRP-HPLC (C₁₈ column, CH₃CN gradient 5-65%, 0.1% TFA, UV analysis 300 nm,28 min) and lyophilization of the collected fractions afforded 6.0 mg(22% yield) of (5) as a white foam. LRMS (ES+) m/z 384.2 (C₁₉H₁₇N₃O₆+Hrequires 384.15); RP-HPLC (300 nm, 28 min run) 15.2 min.

Synthesis of 4′-Trifluoromethoxy-Tolan Dap Hydroxamic Acid Example 18(1S)—N-(2-amino-1-hydroxycarbamoyl-ethyl)-4-(4-trifluoromethoxy-phenylethynyl)-benzamide(5) Preparation of(1S)—N-(2-(Boc)-amino-1-hydroxycarbamoyl-ethyl)-4-ethynyl-benz-amide onHydroxylamine 2-chlorotrityl Resin (3)

Fmoc-Dap resin (1) (1.330 g, 0.931 mmol, 0.70 mmol/g) was swelled in DCM(15 mL) for 2 h and drained. The resin was treated with 20% piperidinein DMF (20 mL) for 1 hour, washed with DMF (3×15 mL) and DCM (3×15 mL)and drained completely. In a separate flask, 4-ethynylbenzoic acid (2)(0.408 g, 2.793 mmol), DIC (0.70 mL, 4.470 mmol), HOBt (0.377 g, 2.793mmol) and DIEA (1.0 mL, 5.7 mmol) were dissolved in DCM (10 mL) and DMF(2 mL), stirred 15 min and added to the resin. After shaking for 36 h,the mixture was drained, washed with DMF (3×15 mL) and DCM (3×15 mL) anddried in vacuo to give 1.290 g of (3) as a yellow resin.

Preparation of(1S)—N-(2-amino-1-hydroxycarbamoyl-ethyl)-4-(4-trifluoromethoxy-phenylethynyl)-benzamide(5)

Resin (3) (120 mg, 0.084 mmol) was swelled in DCM (2 mL) for 1 h anddrained. A solution of 4-(trifluoromethoxy)iodobenzene (4) (96.8 mg,0.336 mmol) and Et₃N (150 μL, 1.10 mmol) in DMF (2.0 mL) was purged witha stream of N₂ bubbles for two minutes and added to the resin. Aftermixing for 5 min, PdCl₂(PPh₃)₂ (18.0 mg, 0.025 mmol) and CuI (8.0 mg,0.042 mmol) were added and the mixture shaken for 24 h. The resin wasdrained, washed with DMF (3×2 mL), DCM (3×2 mL) and cleaved with 10%TFA/DCM (2.0 mL) for 20 min. The solution was collected and the resinwas rinsed with additional 10% TFA/DCM (1.0 mL). The cleavage fractionswere combined, treated with neat TFA (3.0 mL), stirred for 1 h at rt andconcentrated by rotary evaporation to give a crude brown residue.Purification by RP-HPLC (C₁₈ column, CH₃CN gradient 5-55%, 0.1% TFA, UVanalysis 300 nm, 28 min) and lyophilization of the collected fractionsafforded 9.0 mg (25% yield) of (5) as a white solid. LRMS (ES+) m/z408.0 (C₁₉H₁₆F₃N₃O₄+H requires 408.11); RP-HPLC (300 nm, 28 min run)18.0 min.

Example 19 Synthesis ofN-(1-(N-hydroxycarbamoyl)(1S,2R)-2-hydroxypropyl)[4-(4-phenylbuta-1,3-diynyl)phenyl]carboxamide

4-(2,2-Dibromo-vinyl)-benzoic acid methyl ester (2) was made by themethod of Wang, S., et al., J. Org. Chem. 1999, 64, 8873-8879.

A solution of (2) (5.76 g, 18.0 mmol), ethynyl-benzene (3) (2.57 g, 25.2mmol), Pd₂dba₃ (164 mg, 0.18 mmol), tris(4-methoxyphenyl) phosphine(TMPP) (253 mg, 0.72 mmol) were dissolved in argon sparged (5 min.) DMF(60 ml). The reaction was sparged with argon for 1 min. TEA (7.5 ml,54.0 mmol) was added to the stirred reaction mixture that was thenheated under argon at 85° C. for 3.5 hours. The reaction was foundcomplete by LCMS. The reaction was cooled to rt and diluted withEtOAc/hexane (1:1) (300 ml). The organic phase was washed with 1M HCl(2×50 ml), 1M NaOH (3×50 ml), water (2×50 ml), sat. brine (50 ml), driedwith Na₂SO₄, filtered and concentrated under reduced pressure to obtain5.25 g of crude product as an oil. The oil was treated withapproximately 20 ml of a solution of 20% EtOAc/hexane that was heated todissolve the residue. The walls of the flask were washed with the 20%EtOAc/hexane solution (5 ml) that upon cooling gave 1.45 g of pureproduct (31% yield) as a white solid. The balance of the crude reactionproduct was purified by flash chromatography using EtOAc (8%)/hexane aseluant. The pure fractions were evaporated and dried in vacuo to giveaddition product typically 25-30% addition yield.

4-(4-Phenyl-buta-1,3-diynyl)-benzoic acid methyl ester (4) was madeaccording to the method of Wang, S., et al., Org. Lett. 2000, 2(18),2857-2860.

Preparation of 4-(4-Phenyl-buta-1,3-diynyl)-benzoic acid (5)

A 3M aq. solution of NaOH (20 ml) was added to a stirred solution ofmethyl ester (4) (1.45 g, 5.6 mmol) in MeOH (100 ml) at rt. The reactionsolution was heated to reflux for 45 min. until the reaction turnedclear. All of the starting material was gone by TLC and HPLC. Thereaction was cooled to rt and some MeOH (˜50 ml) was removed byevaporation under reduced pressure. Water (100 ml) was added to themixture. Conc. HCl was added dropwise to the stirred solution untilacidic by pH paper (pH2). The white precipitate that formed wascollected by suction filtration. The solid was washed with water (3×20ml) and hexane (2×20 ml) to give after drying 1.35 g of (5) in 99%yield.

Subsequent conversion of compound (5) toN-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)-4-(penta-1,3-diynyl)benzamide(7) was performed according to the method described in Example 12 forthe synthesis ofN-(2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-phenylethynyl-benzamide(compound 5). LCMS MH+363.13.

Example B Synthesis ofN-[(1S)-1-(aminomethyl)-2-(hydroxyamino)-2-oxoethyl]-4-[4-(4-aminophenyl)buta-1,3-diynyl]benzamidePreparation of2-{4-[4-(4-Amino-phenyl)-buta-1,3-diynyl]-benzoylamino}-3-tert-butoxycarbonylamino-propionic acid methyl ester (2)

DIEA (10.5 ml, 60.3 mmol) was added to a stirred solution of4-[4-(4-Amino-phenyl)-buta-1,3-diynyl]-benzoic acid (1) (5.0 g, 19.1mmol), HOBT (2.72 g, 20.1 mmol), EDC (3.85 g, 20.1 mmol) in DMF (80 ml).After 2 min., H-DAP(Boc)-OMe (5.12 g, 2.1 mmol) was added in oneportion. After 12 hours at rt, the reaction was found complete by LCMS.The reaction was diluted with EtOAc/hexane (4:1) (500 ml). The organicphase was washed with 1N NaOH (2×80 ml), water (2×80 ml), sat. brine (80ml), dried with Na₂SO₄, filtered and concentrated under reduced pressureto give crude product. The residue was filtered through a filter plug ofsilica eluting with EtOAc/hexane (4:1). The fractions with product wereevaporated to give 8.02 g of (2) in 91% yield. Subsequent conversion of(2) to the final hydroxamic acid was performed according to the methoddescribed in Example 12 for the synthesis ofN-(2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-phenylethynyl-benzamide(compound 5).

Synthesis of 4-(Buta-1,3-diynyl)-benzoic Acid (4) for making 1,3-diynylanalogues (such as Example 20 below) Preparation of4-(4-trimethylsilanyl-buta-1,3-diynyl)-benzoic acid methyl ester (3)

A solution of methyl 4-iodobenzoate (2) (4.510 g, 17.2 mmol),PdCl₂(PPh₃)₂ (483 mg, 0.690 mmol), and CuI (262 mg, 1.37 mmol) in CH₃CN(50 mL) was cooled to 0° C. under N₂ atmosphere in the absence of light.Triethylamine (7.2 mL, 52.0 mmol) was added, followed bytrimethylsilyl-1,3-butadiyne (1) (5.240 g, 42.8 mmol) and the reactionstirred 3 h at 0° C. and 30 h at ambient temperature. Removal of solventby rotary evaporation afforded a crude black residue that was purifiedby silica gel chromatography (95:5 hexanes/EtOAc) to give 3.450 g (79%yield) of (3) as a brown solid, mp=67-68° C.

Preparation of 4-(buta-1,3-diynyl)-benzoic acid (4)

Potassium hydroxide (3.700 g, 65.9 mmol) was dissolved in H₂O (10 mL)and added to a solution of (3) (3.420 g, 13.5 mmol) in THF (26 mL) inthe absence of light. After stirring 16 h, the reaction was quenchedwith 1.0 M HCl (120 mL) and the resulting precipitate was filtered,washed with 1:1 hexanes/benzene (150 mL) and dried in vacuo to afford2.100 g (91% yield, 98% pure) of (4) as a brown solid, mp>230° C.Although diyne (4) was found to be unstable at room temperature it couldbe stored for several weeks at 0° C. with only small amounts ofdecomposition observed by TLC. R_(f)=0.2 (4:1 Hexanes/EtOAc); HPLC (300nm, 28 min run) 16.0 min; LRMS (ES+) m/z 171.0 (C₁₁H₆O₂+H requires171.04).

Synthesis of a 3′-Nitrophenyl-Diacetylenic-Dap Hydroxamic Acid Example20N-(1-(N-hydroxycarbamoyl)(1S)-2-aminoethyl){4-[4-(3-nitrophenyl)buta-1,3-diynyl]phenyl}carboxamide(6) Preparation of Fmoc-Dap(Boc)-NHOH on hydroxylamine 2-chlorotritylresin (2)

A suspension of N-Fmoc-hydroxylamine 2-chlorotrityl resin (1) (3.288 g,2.53 mmol, 0.77 mmol/g, Novabiochem) in DCM (40 mL) was shaken for 2 hand drained. The resin was treated with 20% piperidine in DMF (40 mL)for 1 hour, washed with DMF (2×40 mL), treated a second time with 20%piperidine in DMF (40 mL), washed with DMF (3×40 mL) and DCM (3×40 mL)and drained completely. In a separate flask, Fmoc-Dap(Boc)-OH (3.175 g,7.44 mmol), HATU (2.829 g, 7.44 mmol) and DIEA (4.3 mL, 24.7 mmol) weredissolved in DMF (35 mL), stirred three minutes and added to the resin.After shaking for 48 h, the mixture was drained, washed with DMF (4×40mL) and DCM (4×40 mL) and dried in vacuo to give 3.530 g of (2) as ayellow resin.

Preparation of(S)—N-(2-N-Fmoc-amino-1-hydroxycarbamoyl-ethyl)-4-buta-1,3-diynyl-benzamideon hydroxylamine 2-chlorotrityl resin (4)

Resin (2) (3.530 g, 2.53 mmol, 0.71 mmol/g) was swelled in DCM (40 mL)for 2 h and drained. The resin was treated with 20% piperidine in DMF(40 mL) for 1 hour, washed with DMF (4×40 mL) and DCM (4×40 mL) anddrained completely. In a separate flask, 4-buta-1,3-diynyl-benzoic acid(3) (1.076 g, 6.32 mmol), EDCI (1.457 g, 7.60 mmol), HOBt (1.048 g, 7.75mmol) and DIEA (2.2 mL, 12.6 mmol) were dissolved in DCM (25 mL) and DMF(5 mL), stirred 45 min and added to the resin. After shaking for 48 h,the mixture was drained, washed with DMF (4×40 mL) and DCM (4×40 mL) anddried in vacuo to give 3.35 g of (4) as a pale brown resin.

Preparation of(S)—N-(2-amino-1-hydroxycarbamoyl-ethyl)-4-[4-(3-nitro-phenyl)-buta-1,3-diynyl]-benzamide(6)

Resin (4) (176 mg, 0.135 mmol) was swelled in DCM (3 mL) for 1 h anddrained. A solution of 1-iodo-3-nitrobenzene (5) (118 mg, 0.474 mmol)and Et₃N (200 μL, 1.43 mmol) in DMF (3.0 mL) was purged with a stream ofN₂ bubbles for two minutes and added to the resin. After mixing for 5min, PdCl₂(PPh₃)₂ (6.0 mg, 0.009 mmol) and CuI (10.0 mg, 0.052 mmol)were added and the mixture shaken for 36 h. The resin was drained,washed with DMF (4×3 mL), DCM (4×3 mL) and cleaved with 10% TFA/DCM (2mL) for 20 min. The solution was collected and the resin was rinsed withadditional 10% TFA/DCM (2 mL). The cleavage fractions were combined,treated with neat TFA (4.0 mL), stirred for 1 h at rt and concentratedby rotary evaporation to give a crude brown residue. Purification byRP-HPLC (C₁₈ column, CH₃CN gradient 5-65%, 0.1% TFA, UV analysis 300 nm,30 min) and lyophilization of the collected fractions afforded 12.0 mg(22%) of 470 as a white solid. LRMS (ES+) m/z 392.9 (C₂₀H₁₆N₄O₅+Hrequires 393.11); RP-HPLC (300 nm, 30 min run) 14.9 min.

Synthesis of 4′-Benzamide Diacetylene Dap Hydroxamic Acid Example 21N-((2S)-amino-1-hydroxycarbamoyl- ethyl)-4-{4-[4-(2-amino-ethylcarbamoyl)-phenyl]-buta-1,3-diynyl}-benzamide (3)

(1S)—N-(2-(Boc)-amino-1-hydroxycarbamoyl-ethyl)-4-ethynyl-benz-amide onhydroxylamine-2-chloro resin (1) (145 mg, 0.111 mmol) was swelled in DCM(2 mL) for 1 h and drained. A solution of 4-ethynylbenzamide (2) (124mg, 0.288 mmol) and Et₃N (100 μL, 0.72 mmol) in DMF (2.0 mL) was addedand the resin agitated for 5 min. A mixture of PdCl₂(PPh₃)₂ (21 mg,0.030 mmol) and CuI (22 mg, 0.110 mmol) was added and the resin wasagitated for 60 h. The resin was drained, washed with DMF (3×2 mL), DCM(3×2 mL) and cleaved with 10% TFA/DCM (1.5 mL) for 20 min. The solutionwas collected and the resin was rinsed with additional 10% TFA/DCM (1.0mL). The cleavage fractions were combined, treated with neat TFA (2.0mL), stirred for 1 h at rt and concentrated by rotary evaporation togive a crude brown residue. Purification by RP-HPLC (C₁₈ column, CH₃CNgradient 5-55%, 0.1% TFA, UV analysis 300 nm, 26 min) and lyophilizationof the collected fractions afforded 2.6 mg (5% yield) of (3). LRMS (ES+)m/z 434.0 (C₂₃H₂₃N₅O₄+H requires 434.19); RP-HPLC (300 nm, 26 min run)15.3 min.

Synthesis of N-[4-Butadiynyl-benzoyl]-Thr(tBu) on Resin (Continued tomake Examples 22 and 23) Preparation of (2S,3R)-2-N-Fmoc-amino-3-tert-butoxy-N-hydroxy-butyramide on hydroxylamine2-chlorotrityl resin (2)

A suspension of N-Fmoc-hydroxylamine 2-chlorotrityl resin (1) (3.188 g,2.45 mmol, 0.77 mmol/g, Novabiochem) in DCM (40 mL) was shaken for 2 hand drained. The resin was treated with 20% piperidine in DMF (40 mL)for 1 hour, washed with DMF (2×40 mL), treated a second time with 20%piperidine in DMF (40 mL), washed with DMF (3×40 mL) and DCM (3×40 mL)and drained completely. In a separate flask, Fmoc-Thr(tBu)-OH (2.927 g,7.36 mmol), HATU (2.798 g, 7.36 mmol) and DIEA (4.3 mL, 24.6 mmol) weredissolved in DMF (40 mL), stirred three minutes and added to the resin.After shaking for 24 h, the mixture was drained, washed with DMF (4×40mL) and DCM (4×40 mL) and dried in vacuo to give 3.500 g of (2) as ayellow resin.

Preparation of4-buta-1,3-diynyl-N-(2-tert-butoxy-1-hydroxycarbamoyl-propyl)-benzamideon hydroxylamine 2-chlorotrityl Resin (4)

Resin (2) (2.030 g, 1.56 mmol, 0.77 mmol/g) was swelled in DCM (20 mL)for 2 h and drained. The resin was treated with 20% piperidine in DMF(20 mL) for 1 hour, washed with DMF (4×20 mL) and DCM (4×20 mL) anddrained completely. In a separate flask, 4-buta-1,3-diynyl-benzoic acid(3) (0.617 g, 3.63 mmol), EDCI (0.834 g, 4.35 mmol), HOBt (0.588 g, 4.35mmol) and DIEA (1.0 mL, 5.7 mmol) were dissolved in DCM (15 mL) and DMF(4 mL), stirred 45 min and added to the resin. After shaking for 36 h,the mixture was drained, washed with DMF (4×20 mL) and DCM (4×20 mL) anddried in vacuo to give 1.900 g of (4) as a pale brown resin.

Synthesis of Diacetylenic Threonine Hydroxamic Acids Example 22(2S,3R)-4-[4-(3-aminomethyl-phenyl)-buta-1,3-diynyl]-N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide(3)

Resin (1) (resin (4) obtained from previous synthesis) (100 mg, 0.077mmol) was swelled in DCM (2 mL) for 1 h and drained. A solution of3-iodobenzylamine hydrochloride (2) (83.0 mg, 0.308 mmol) and Et₃N (250μL, 1.80 mmol) in DMF (1.5 mL) was purged with a stream of N₂ bubblesfor two minutes and added to the resin. After mixing for 5 min,PdCl₂(PPh₃)₂ (11.0 mg, 0.016 mmol) and CuI (7.0 mg 0.037 mmol) wereadded and the mixture shaken for 36 h. The resin was drained, washedwith DMF (4×2 mL), DCM (4×2 mL) and cleaved with 10% TFA/DCM (1.5 mL)for 20 min. The solution was collected and the resin was rinsed withadditional 10% TFA/DCM (1.5 mL). The cleavage fractions were combined,treated with neat TFA (3.0 mL), stirred for 1 h at rt and concentratedby rotary evaporation to give a crude brown residue. Purification byRP-HPLC (C₁₈ column, CH₃CN gradient 5-65%, 0.1% TFA, UV analysis 300 nm,28 min) and lyophilization of the collected fractions afforded 4.3 mg(14%) of (3) as a white solid. LRMS (ES+) m/z 392.0 (C₂₂H₂₁N₃O₄+Hrequires 392.15); RP-HPLC (300 nm, 28 min run) 10.0 min.

Synthesis of Diacetylenic Benzylamine Analogues Example 23

(1S,2R)—N-2-bydroxy-1-hydroxycarbamoyl-propyl)-4-[4-(4-meorphoin-4-ylmethyl-phenyl)-buta-1,3-diynyl]-benzamide(4)

Preparation of Threonine Diacetylenic Benzaldehyde on Resin (3)

Resin (1) (resin (4) obtained from prior synthesis)) (1.00 g, 0.77 mmol)was pre-swelled in DCM (25 mL) for 14 h and drained. A solution of4-iodobenzaldehyde (2) (715 mg, 3.08 mmol) and Et₃N (1.00 mL, 7.17 mmol)in DMF (20 mL) was purged with N₂ for two minutes and added to theresin. After mixing for 5 min, PdCl₂(PPh₃)₂ (40.0 mg, 0.057 mmol) andCuI (19.0 mg, 0.100 mmol) were added and the reaction shaken for 48 h.The resin was drained, washed with DMF (4×20 mL), DCM (4×20 mL) anddried in vacuo to give 1.100 g of (3) as a dark yellow resin.

Preparation of (1 S.2R)—N-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-[4-(4-morpholin-4-ylmethyl-phenyl)-buta-1,3-diynyl]-benzamide(4)

A solution of morpholine (75 μL, 0.860 mmol) and trimethyl orthoformate(100 μL, 0.914 mmol) in THF (3.0 mL) was added to a Teflon-linedscrew-capped vial containing the resin-bound diacetylenic benzaldehyde(3). The resin was agitated for 10 min, treated successively with aceticacid (100 μL, 1.75 mmol) and a solution of NaCNBH₃ (40.0 mg, 0.637nmmol) in MeOH (1.0 mL) and shaken for 44 h. The resin was filtered,washed with DMF (3×3 mL) and DCM (3×3 mL) and drained. Cleavage from theresin was achieved by treatment with 10% TFA/DCM (2.0 mL) and shaking 20min. The solution was collected and the resin was rinsed with additional10% TFA/DCM (2.0 mL). The cleavage fractions were combined, treated withneat TFA (3.0 mL), stirred for 1 h at rt and concentrated by rotaryevaporation to give a crude yellow residue. Purification by RP-HPLC (C₁₈column, CH₃CN gradient 5-35%, 0.1% TFA, UV analysis 300 nm, 18 min) andlyophilization of the collected fractions afforded 19.0 mg (29%) of (4)as a fluffy yellow solid. LRMS (ES+) m/z 462.0 (C₂₆H₂₇N₃O₅+H requires462.10); HPLC (300 nm, 18 min run) 10.3 min.

Synthesis of 4′-Benzamide Diacetylene Threonine Hydroxamic Acid Example24(1S,2R)—N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-4-{4-[4-(2-amino-ethylcarbamoyl)-phenyl]-buta-1,3-diynyl}-benzamide(5) Preparation of N-(2-trityl-amino-ethyl)-4-ethynyl-benzamide (3)

To a solution of 4-ethynylbenzoic acid (1) (292 mg, 2.00 mmol), EDCI(382 mg, 2.00 mmol), and HOBt (270 mg, 2.00 mmol) in DMF (10 mL) wasadded N-trityl ethylenediamine (2) (810 mg, 2.67 mmol) and DIEA (1.4 mL,8.0 mmol). The reaction mixture was stirred 24 h, diluted with EtOAc(200 mL), washed with 0.5 M HCl (60 mL), saturated NaHCO₃ (2×60 mL), H₂O(4×60 mL), dried over MgSO₄ and concentrated to give 836 mg (97% yield)of (3) as a white solid, mp 50-51° C. R_(f)=0.40 (1:1 Hexanes/EtOAc).

Preparation of(1S,2R)—N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-4-{4-[4-(2-amino-ethylcarbamoyl-phenyl]-buta-1,3-diynyl}-benzamide(5)

Resin (4) (resin (1) from above syntheses) (150 mg, 0.116 mmol) wasswelled in DCM (2 mL) for 1 h and drained. A solution of4-ethynylbenzamide (3) (151 mg, 0.350 mmol) and Et₃N (150 μL, 1.10 mmol)in DMF (2.0 mL) was added and the resin agitated for 5 min. A mixture ofPdCl₂(PPh₃)₂ (21 mg, 0.030 mmol) and CuI (28 mg, 0.147 mmol) was addedand the resin was agitated for 60 h. The resin was drained, washed withDMF (3×2 mL), DCM (3×2 mIL) and cleaved with 10% TFA/DCM (1.5 mL) for 20min. The solution was collected and the resin was rinsed with additional10% TFA/DCM (1.0 mL). The cleavage fractions were combined, treated withneat TFA (2.0 mL), stirred for 1 h at rt and concentrated by rotaryevaporation to give a crude brown residue. Purification by RP-HPLC (C₁₈column, CH₃CN gradient 5-65%, 0.1% TFA, UV analysis 300 nm, 26 min) andlyophilization of the collected fractions afforded 2.0 mg (4% yield) of(5). LRMS (ES+) m/z 449.1 (C₂₄H₂₄N₄O₅+H requires 449.18); RP-HPLC (300nmn, 26 min run) 17.0 min.

Synthesis of 3′-Pyridine Diacetylene Threonine Hydroxamic Acid Example25N-((2R)-hydroxy-(1S)-hydroxycarbamoyl-propyl)-4-(4-pyridin-3-yl-buta-1,3-diynyl)-benzamide(3)

(1S,2R)—N-(2-tert-butoxy-1-hydroxycarbamoyl-propyl)-4-ethynyl-benzamideon hydroxylamine 2-chlorotrityl resin (1) (142 mg, 0.109 mmol) wasswelled in DCM (2 mL) for 1 h and drained. A solution of3-ethynylpyridine (2) (38 mg, 0.368 mmol) and Et₃N (200 μL, 1.4 mmol) inDMF (2 mL) was added and the resin agitated for 5 min. A mixture ofPdCl₂(PPh₃)₂ (22 mg, 0.031 mmol) and CuI (25 mg, 0.131 mmol) was addedand the resin was agitated for 72 h. The resin was drained, washed withDMF (3×2 mL), DCM (3×2 mL) and cleaved with 10% TFA/DCM (1.5 mL) for 20min. The solution was collected and the resin was rinsed with additional10% TFA/DCM (1.0 mL). The cleavage fractions were combined, treated withneat TFA (2.0 mL), stirred for 1 h at rt and concentrated by rotaryevaporation to give a crude brown residue. Purification by RP-HPLC (C₁₈column, CH₃CN gradient 5-65%, 0.1% TFA, UV analysis 300 nm, 24 min) andlyophilization of the collected fractions afforded 4.4 mg (11% yield) of(3). LRMS (ES+) m/z 364.0 (C₂₀H₁₇N₃O₄+H requires 364.13); RP-HPLC (300nm, 24 min run) 11.2 min.

Example 26 Synthesis of N-(1-(N-hydroxycarbamoyl)(1S,2R)-2-hydroxypropyl){4-[4-(6-morpholin4-yl(3-pyridyl))buta-1,3-diynyl]phenyl}carboxamide (5)Preparation of 4-[4-(6-Chloro-pyridin-3-yl)-buta-1,3-diynyl]-benzoicacid methyl ester

4-[4-(6-Chloro-pyridin-3-yl)-buta-1,3-diynyl]-benzoic acid was madeaccording to the method of Wang Shen and Sheela A. Thomas in Org. Lett.2000, 2(18), 2857-2860.

A solution of 4-(2,2-dibromo-vinyl)-benzoic acid methyl ester (1) (9.6g, 30.0 mmol), 2-chloro-5-ethynyl-pyridine (2) (5.43 g, 39.0 mmol),Pd₂dba₃ (274 mg, 0.3 mmol), tris(4-methoxyphenyl) phosphine (TMPP) (422mg, 1.2 mmol) were dissolved in argon sparged (5 min.) DMF (60 ml). Thereaction was sparged with argon for 1 min. TEA (12.5 ml, 90.0 mmol) wasadded to the stirred reaction mixture that was then heated under argonat 85° C. for 3 hours. The reaction was found complete by LCMS. Thereaction was cooled to rt and diluted with EtOAc/hexane (1:1) (500 ml).The organic phase was washed with 1 M NaOH (2×80 ml), water (2×80 ml),sat. brine (80 ml), dried with Na₂SO₄, filtered concentrated underreduced pressure to give crude product. The residue was filtered througha filter plug of silica eluting with EtOAc/hexane (1:1). The fractionswith product were evaporated to give 9.06 g of product in good purity(˜96% pure). The material was taken on without further purification.

Preparation of 4-[4-(6-Chloro-pyridin-3-yl)-buta-1,3-diynyl]-benzoicAcid (3)

A 6M aq. solution of NaOH (15 ml) was added to a stirred solution of4-[4-(6-Chloro-pyridin-3-yl)-buta-1,3-diynyl]-benzoic acid methyl ester(9.06 g, 30 mmol) in MeOH (350 ml) at rt. The reaction solution washeated to reflux for 3 hours. The reaction stayed a mixture and did notturn clear. HPLC and LCMS indicated that the reaction was forming sideproducts. The reaction was cooled to rt and some MeOH (˜200 ml) wasremoved by evaporation under reduced pressure. Water (400 ml) was addedto the mixture. Conc. HCl was added dropwise to the stirred solutionuntil acidic by pH paper (pH2). The yellow precipitate that formed wascollected by suction filtration. The solid was washed with water (3×20ml) and hexane (2×20 ml) to give the crude product. HPLC indicated thatthere was approximately 40% product in the mixture. The crude reactionproduct was purified by flash chromatography using EtOAc (8-10%)/hexaneas eluant. The pure fractions were evaporated and dried in vacuo to give4.2 g of product (3) in 50% yield.

Preparation of[4-[4-(6-chloro-pyridin-3-yl)-buta-1,3-diynyl]-benzoyl]-HN-Thr(OtBu)-hydroxamicacid trityl resin (4)

(3) was coupled to a tert-butyl protected threonine pre-loaded onhydroxylamine 2-chlorotrityl resin following the same procedure as usedfor Example 26. The coupling employed DIC and HOBT.[N-Fmoc-hydroxylamine 2-chlorotrityl resin was purchased fromNovabiochem cat. #01-64-0165.]

Preparation ofN-(2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-[4-(6-morpholin-4-yl-pyridin-3-yl)-buta-1,3-diynyl]-benzamide(5)

A solution of morpholine (300 uL) in NMP (1 ml) was added to a vialcontaining (4) (150 mg, 0.12 mmol). The reaction mixture was purged withargon and heated to 85-90° C. for 24 hours. The resin was drained andwashed with DMF and DCM alternately several times. The product wascleaved from the resin through treatment with a TFA/water solution(80:20) (1.5 ml) for 45 min. The resin was filtered and washed withfresh TFA/water solution (80:20) (0.5 ml). The combined TFA and organicfractions were diluted with CH₃CN/water (1:1) (10 ml), water (2 ml) andlyophilized. The crude product was purified by prep. HPLC. The crudeproduct was dissolved in DMSO (1 ml), passed through a Teflon syringefilter, and the clear filtrate was injected on a preparative HPLC. Thepurification used a 20×50 mm Ultro 120 C18 column running a 22 ml/min 2%gradient (AcCN/water, 0.1% TFA) for 16 min. The purified fractions werelyophilized to dryness to give 2.2 mg of pure product (5) as the TFAsalt (˜32% yield).

Example 27 Synthesis of4-[4-(4-Amino-phenyl)-buta-1,3-diynyl]-N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide(4) Preparation of 2{4-[4-(4-Amino-phenyl)-buta-1,3-diynyl]-benzoylamino}-3-tert-butoxycarbonyloxy-butyric hydroxamic acid trityl resin (3)

DIEA (2.7 ml, 15.6 mmol) was added to a stirred solution of4-[4-(4-Amino-phenyl)-buta-1,3-diynyl]-benzoic acid (2) (1.64 g, 6.3mmol), HOBT (0.85 g, 6.3 mmol), DIC (0.98 ml, 6.3 mmol) in DMF (50 ml).After 2 min., H-Thr(Boc)-NHO-Trt resin (1) (5.8 g, 4.5 mmol) was addedin one portion. [N-Fmoc-hydroxylamine 2-chiorotrityl resin was purchasedfrom Novabiochem cat. #01-64-0165.] After 12 hours at rt, the reactionwas found complete by LCMS. The resin was drained and washed with DMFand DCM alternately 3 times each. The product (3) was used as is insubsequent reactions without further treatment.

Preparation of4-]4-(4-Ainino-phenyl)-buta-1,3-diynyl]-N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide (4)

The product (4) (120 mg, 0.09 mmol) was cleaved from resin (3) throughtreatment with a TFA/water solution (80:20) (1.5 ml) for 45 min. Theresin was filtered and washed with fresh TFA/water solution (80:20) (0.5ml). The combined TFA and organic fractions were diluted withCH₃CN/water (1:1) (10 ml), water (2 ml) and lyophilized. The crudeproduct was purified by prep. HPLC. The crude product was dissolved inDMSO (1 ml), passed through a Teflon syringe filter, and the clearfiltrate was injected on a preparative HPLC. The purification used a20×50 mm Ultro 120 C18 column running a 22 ml/min 2% gradient(AcCN/water, 0.1% TFA) for 16 min. The purified fractions werelyophilized to dryness to give 2.2 mg of pure product as the TFA salt.The product was lyophilized again from CH₃CN/water with 10 equivalentsof HCl to remove most of the TFA to yield 2 mg of (4) as the HCl salt(˜53% yield).

Example 28 Synthesis of4-{4-[4-(2-Dimethylamino-acetylamino)-phenyl]-buta-1,3-diynyl}-N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide(6) (Continued from Compound (3) of Example 27 Above) Preparation of2-{4-(4-[4-(2-Bromo-acetylamino)-phenyl]-buta-1,3-diynyl}-benzoylamino)-3-tert-butoxycarbonyloxy-butyricacid hydroxamate trityl resin (5)

A solution of bromo-acetyl chloride (0.75 g, 0.58 mmol) in DCM (2 ml)was added to a mixture of (3) (0.75 g, 0.58 mmol), lutidine (1.1 ml, 9.2mmol) and DCM (4 ml) at rt with shaking. After shaking for 1.5 hours,the reaction was found complete by LCMS. The resin was drained andwashed with DCM (2×10 ml), DMF (3×10 ml) and DCM (3×10 ml) again. Theresin was drained and dried in vacuo. The product resin (5) was used asis in subsequent reactions without further treatment.

Preparation of4-{4-[4-(2-Dimethylamino-acetylamino)-phenyl]-buta-1,3-diynyl}-N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide(6)

A solution of dimethyl amine (0.2 ml) in NMP (1.2 ml) was added to (5)(125 mg, 0.09 mmol) at rt with shaking. After shaking for 12 hours, thereaction was found complete by LCMS. The resin was drained and washedwith DCM (2×10), DMF (3×10) and DCM (3×10) again. The product (6 ml) wascleaved from the resin through treatment with a TFA/water solution(80:20) (1.5 ml) for 45 min. The resin was filtered and washed withfresh TFA/water solution (80:20) (0.5 ml). The combined TFA and organicfractions were diluted with CH₃CN/water (1:1) (10 ml), water (2 ml) andlyophilized. The crude product was purified by prep. HPLC. The crudeproduct was dissolved in DMSO (1 ml), passed through a Teflon syringefilter, and the clear filtrate was injected on a preparative HPLC. Thepurification used a 20×50 mm Ultro 120 C18 column running a 22 ml/min 2%gradient (AcCN/water, 0.1% TFA) for 16 min. The purified fractions werelyophilized to dryness to give 2 mg of pure product (6) as the TFA salt(˜37% yield).

Example 29 Synthesis of4-{4-[4-(2-Amino-4-methyl-pentanoylamino)-phenyl]-buta-1,3-diynyl}-N-(2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide(7) (Continued from compound (3) of Example 27 Above)

A solution of Fmoc-L-leucine (0.135 g, 0.38 mmol), HATU (0.146 g, 0.38mmol) in DMF (1.5 ml) was made. After 2 min. of shaking, the activatedacid was added to (3) (125 mg, 0.09 mmol) at rt with shaking. Aftershaking for 36 hours, the reaction was drained and washed with DCM (2×4ml), DMF (3×4 ml) and DCM (3×4 ml) again. The resin was treated with 20%piperizine in DMF (4 ml) for 2 hours twice. The resin was drained andwashed with DMF and DCM alternately several times. The product wascleaved from the resin through treatment with a TFA/water solution(80:20) (1.5 ml) for 45 min. The resin was filtered and washed withfresh TFA/water solution (80:20) (0.5 ml). The combined TFA and organicfractions were diluted with CH3CN/water (1:1) (10 ml), water (2 ml) andlyophilized. The crude product was purified by prep. HPLC. The crudeproduct was dissolved in DMSO (1 ml), passed through a Teflon syringefilter, and the clear filtrate was injected on a preparative HPLC. Thepurification used a 20×50 mm Ultro 120 C18 column running a 22 ml/min 2%gradient (AcCN/water, 0.1% TFA) for 16 min. The purified fractions werelyophilized to dryness to give 1.7 mg of pure product (7) as the TFAsalt (˜30% yield).

Example 30 Step 1

NaBH(OAc)₃ (1.38 g, 6.03 mmol) was added at 0° C. to a solution of4-iodobenzylaldehyde (1.0 g, 4.31 mmol) and morpholine (462 mg, 5.37mmol) in THF (30 mL). The reaction mixture was then warmed to rt andstirred overnight. The solvent was removed (RV) and the residue wasextracted with EtOAc (2×) from aqueous NaHCO₃ (pH=8˜9). The combinedorganic extracts were dried (NaSO₄) and concentrated to dryness (RV).Column chromatography (silica gel, EtOAc/DCM 0-100%) yielded compound(1) (980 mg, 74.8% yield, M+=304.0).

Step 2

A solution of compound (1) (2.0 g, 6.6 mmol) and propargyl alcohol (407mg, 7.26 mmol) in THF (20 mL) was purged by nitrogen for 30 min,followed by addition of PdCl₂(PPh₃)₂ (55.6 mg, 0.079 mmol) and CuI (30mg, 0.158 mmol). The reaction mixture was cooled to 0° C. and triethylamine (1.0 g, 9.9 mmol) was added. The reaction mixture was warmed to rtand stirred overnight under N₂. The reaction mixture was filteredthrough a plug of celit and the filtrate was concentrated. The residuewas treated with water (200 mL) and extracted with EtOAc (200 mL×2). Thecombined organic layers were washed with water (200 mL×2), brine (200mL) and dried (NaSO₄). The crude product was purified by chromatographyon silica gel eluting with EtOAc/Hexane (0-50%) to give 1.5 g of theproduct (2) (98.7% yield, M+H⁺=232.0).

Step 3

Dess-Martin reagent (1.37 g, 3.25 mmol) was added to a solution ofcompound (2) (500 mg, 2.1 mmol) in CH₂Cl₂ (5 mL) at 0° C. The reactionmixture was warmed to rt and stirred overnight. The reaction mixture wasdiluted with CH₂Cl₂ (200 mL), basified with NaHCO₃ (saturated) to pH=8-9and then washed with water (200 mL) and brine (200 mL), dried (NaSO₄)and concentrated under reduced pressure. The crude product (3) (900 mg,MS+H⁺=230.0) was not very stable and used directly in the next reactionwithout further purification.

Step 4

A mixture of 4-bromomethyl-benzoylic methyl ester (1.0 g, 4.37 mmol) andP(OEt)₃ (3 mL) in a sealed tube was heated at 150° C. overnight. Cooledto rt, the reaction mixture was diluted with EtOAc (200 mL), washed withwater (100 mL×2) and brine (100 mL), dried (NaSO₄) and concentratedunder reduced pressure. The crude product was purified by chromatographyon silica gel eluting with EtOAc/Hexane (0-100%) to give compound (4)(800 mg, 64.0% yield, M+H⁺=286.9).

Step 5

To a solution of compound (4) (741 mg, 2.59 mmol) in THF (5 mL) wasadded NaH (114 mg, 2.85 mmol). Stirred at rt for one hour, followed byaddition of compound (3) (900 mg) in THF solution (5 mL), the resultingmixture was stirred at rt overnight and concentrated under reducedpressure. The residue was treated with EtOAc (300 mL), washed with water(200 ml×2) and brine (200 ml), dried (NaSO₄) and concentrated underreduced pressure. The crude product was purified by chromatography onsilica gel eluting with EtOAc/Hexane (0-60%) to give compound (5) (188mg, 20.0% yield, M+H⁺=362.0).

Step 6

To a solution of compound (5) (188 mg, 0.521 mmol) in MeOH (5 mL) wasadded NaOH (84 mg, 1.08 mmol). The reaction mixture was heated at refluxfor 4 hrs and cooled to rt, the reaction mixture was neutralized to pH=7with HCl solution (4 N in dioxane). Removal of solvent under reducedpressure, the solid residue was washed with CH₂Cl₂. The combined CH₂Cl₂was evaporated to give crude compound (6) (259 mg, 70% purity, 100%yield, MS⁺=348.0).

Step 7

To a solution of compound (5) (320 mg, 0.886 mmol) in MeOH (5 mL) wasadded NaOH (142 mg, 3.54 mmol). The reaction mixture was heated atreflux for 4 hrs and cooled to rt. The reaction mixture was concentratedunder reduced pressure. The crude product was purified by prep. HPLC togive compound (6a) (105.5 mg, 25.8% yield, MS⁺=348.0) and (6b) (30.9 mg,7.4%, MS⁺=348.0) as TFA salt.

Step 8

To a solution of compound (6) (44 mg, 0.126 mmol) and methyl 3-Boc-2,3-diaminopropanoade (64 mg, 0.253 mmol) and PyBOP (131 mg, 0.253 mmol)in DMF (2 mL) was added DIPEA (81 mg, 0.65 mmol). The reaction mixturewas stirred at rt. overnight and diluted with water (50 ml), extractedwith EtOAc (50 ml×2). The combined organic layers were washed with water(50 ml×2), brine (50 ml), dried (NaSO₄) and concentrated under reducedpressure. The crude product was purified by chromatography on silica geleluting with MeOH/CH₂Cl₂ (0-5%) to give product 7 (47 mg, 68.1%,MS⁺=548.1).

Step 9

To a solution of compound (7) (48 mg, 0.08 mmol) in CH₂Cl₂ (1 mL)/MeOH(2 mL) was added NH₂OH.HCl (61 mg, 0.88 mmol) and NaOH (58 mg, 1.05mmol). The reaction mixture was stirred at rt for 3 days, and thendiluted with CH₂Cl₂ (20 mL), then treated with NH₄Cl (saturated)solution to adjust pH=7. The organic layer was separated, and the aq.layer was extracted with CH₂Cl₂ (20 mL×2). The combined CH₂Cl₂ layerswere washed with brine (20 ml), dried (NaSO₄) and concentrated underreduced pressure. The crude product was purified by chromatography onsilica gel eluting with CH₂Cl₂₁MeOH (0-10%) to give product (8) (10 mg,22.8%. MS⁺=549.1)

Step 10

The desired product (9a) (90 mg, 89.7%, MS⁺=463.1) was prepared by thereaction of compound (6a) (100 mg, 0.217 mmol), L-Threonine ethyl ester(75 mg, 0.434 mmol), HATU (165 mg, 0.434 mmol) and DIPEA (138 mg, 1.07mmol) in DMF (3 mL) according to the synthetic procedure for thepreparation of compound (7).

Step 11

The desired product (9b) (26 mg, 90%, MS⁺=463.1) was prepared by thereaction of compound (6a) (30 mg, 0.065 mmol), L-Threonine ethyl ester(22 mg, 0.13 mmol), HATU (49 mg, 0.138 mmol) and DIPEA (42 mg, 0.325mmol) in DMF (1 mL) according to the synthetic procedure for thepreparation of compound (7).

Step 12

The desired product (10) (36 mg, 46.0%, MS⁺=461.1) was prepared by thereaction of compound (6) (60 mg, 0.17 mmol), valine methyl ester (54 mg,0.323 mmol), HATU (128 mg, 0.338 mmol) and DIPEA (104 mg, 0.81 mmol) inDMF (3 mL) according to the synthetic procedure for the preparation ofcompound (7).

Step 13

To a solution of compound (8) (48 mg, 0.088 mmol) in CH₂Cl₂ (1 ml) wasadded HCl (1 ml, 4 N in dioxane). The reaction mixture was stirred atrt. for 2 hrs. and concentrated under reduced pressure. The solidresidue was purified by prep. HPLC to give compound (30-11a) (53 mg, 90%yield) and (30-11b) (0.3 mg, 0.5% yield) as double TFA salt.

Step 14

The desired product (30-12a) (53.8 mg, 14.9%, MS⁺=464.1) was prepared bythe reaction of compound (9a) (90 mg, 0.195 mmol) and NH₂OH.HCl (134 mg,1.95 mmol) and NaOMe (128.7 mg, 2.34 mmol) in MeOH (5 ml) according tothe synthetic procedure for the preparation of compound (8).

Step 15

Compound (30-12b) (6.2 mg, 24% yield) was prepared by the reaction ofcompound (9b) (26 mg, 0.056 mmol) and NH₂OH.HCl (38.7 mg, 0.56 mmol) andNaOMe (37 mg, 0.672 mmol) in MeOH (3 ml) according to the syntheticprocedure for the preparation of compound (8).

Step 16

The desired product (13) is prepared by the reaction of compound (10),NH₂OH.HCl and NaOH in CH₂Cl₂/MeOH according to the synthetic procedurefor the preparation of compound (8).

Step 17

The desired product (14) is prepared by the reaction of compound (6) and(2S,3R)-2-amino-3-Fmoc-butyric acid methyl ester hydrochloride accordingto the synthetic procedure for the preparation of compound (7). Thedesired product (15) is prepared by the reaction of compound (14),NH₂OH.HCl and NaOMe in CH₂Cl₂/MeOH according to the synthetic proceduresfor the preparation of compound (8).

Synthesis of (2S,3R)-2-amino-3-Fmoc-butyric acid methyl esterhydrochloride

(2S,3S)-methyl-3-hydroxy-2-(tritylamino)butanoate (14-2)

To a solution of H-allo-Thr-OMe.HCl (compound 14-1) (CAS 79617-27-9,Chem-Impex, 5 g, 29.5 mmoL) in CH₂Cl₂ (30 mL) and DIEA (15 mL, 88.5mmol) was added trityl bromide (10 g, 32.4 mmol) in CHCl₃ (30 mL)dropwise through an addition funnel at 0° C. The reaction mixture waswarmed to rt slowly and stirred overnight, and washed with water (2×10mL) and brine, dried (Na₂SO₄), filtered and concentrated to give thedesired product as a thick oil, which was solidified to a whitecrystalline material in vacuo. The solid was treated with ethyl etherand sonicated for 30 min, filtered and washed with ethyl ether to givethe pure product (14-2) (8 g, 21.3 mmol, 72% yield, mw 375.5) as a whitesolid.

(2S,3R)-methyl 3-azido-2-(tritylamino)butanoate (14-3)

To a mixture of compound (14-2) (6 g, 16 mmol), PPh₃ (4.2 g, 16 mmol) inCH₂Cl₂ (30 mL) at 0° C. with stirring under nitrogen was added diethylazodicarboxylate (DIAD) (5.2 g, 25.6 mmol) in CH₂Cl₂ (5 mL) slowly,followed by diphenylphosphonic azide (DPPA) in CH₂Cl₂ (5 mL). Thereaction mixture was allowed to warm to rt. After stirring at rtovernight, the reaction was concentrated under reduced pressure. Theresidue was purified by chromatography on silica gel on an ISCO systemeluting with hexane/EtOAc to give the desired product (14-3) (2.9 g,˜7.2 mmol, 45% mw 400.5) as a yellow oil (NMR shows there is DIAD in theproduct).

(2S,3R)-methyl 2-amino-3-azidobutanoate (14-4)

A mixture of compound (14-3) (2.9 g) in CH₂Cl₂ (10 mL) and HCl (5 mL, 4M in dioxane) was stirred overnight. The reaction mixture wasconcentrated and the residue was treated with hexane. The suspensionformed was filtered to give the desired product (14-4) (600 mg, 52%) asa white solid. (Alternately, compound (14-3) can be dissolved in THF (1g: 8 mL), HCl 2M (3 eq.) in ether can be added at 0° C. The reaction isallowed to warm and is complete after ˜3 h. The reaction mixture can bediluted with ether to precipitate the product as an HCl salt which isfiltered and washed with ether. The resulting white powder, after dryingin vacuo, is highly pure and gives about 86% yield, mw 158.2 free baseand 194.2 with 1 HCl salt. No further purification is needed.)

(2S,3R)-3-Azido-2-Boc-amino-butyric acid methyl ester (14-5)

A solution of di-tert-butyl dicarbonate (810 mg, 3.7 mmol),(2S,3R)-2-amino-3-azido-butyric acid methylester hydrochloride (compound14-4) (660 mg, 3.38 mmol) and DIEA (645 μl, 3.7 mmol) in i-PrOH (10 ml)was maintained at ambient temperature for 10 h. The reaction mixture wasevaporated in vacuum. The residue was dissolved in water (4 mL) andextracted with hexane/ether (1:1) (4×10 mL). The combined organic layerswere dried (Na₂SO₄) and concentrated under reduced pressure. The residuewas subjected to flash chromatography on CombiFlash® Companion unitequipped with RediSep® flash column (normal phase, 35-60 micron averageparticle size silicagel, 40 g, Teledyne Isco); flow rate=35 ml/min;injection volume 2 ml (EtOAc); mobile phase A: hexane; mobile phase B:EtOAc; gradient 0-50% B in 1 h. Fractions containing the desired productwere combined and concentrated under reduced pressure to providecompound (14-5) (564 mg, 2.2 mmol, 65% yield, mw 258.3) as colorlessoil. (Alternately, the reaction can be run using 1.2 eq Boc₂O and 2.5 eqDIEA in ACN (1 g (14-5) in 30 mL ACN). After stirring overnight, thereaction was concentrated and purified by silica chromatography elutingwith 10-15% EtOAc/Hexane. The product fractions were collected to give98% yield, >98% purity as a clear oil.

(2S,3R)-3-Amino-2-tert-butoxycarbonylamino-butyric acid methyl ester(14-6)

Compound (14-5) (786 mg, 3.05 mmol) was dissolved in methanol (20 ml)followed by the addition of Pd/C (5% wt, 200 mg). The reaction mixturewas subjected to hydrogenation (Parr apparatus, 80 psi) at ambienttemperature for 40 min. The solid catalyst was filtered and washed withmethanol. The filtrate was evaporated under reduced pressure to providecompound (14-6) (682 mg, 96% yield, mw 232.3) as colorless oil.

(2S,3R)-2-Boc-amino-3-Fmoc-aminobutyric acid methyl ester (14-7)

A mixture of compound (14-6) (682 mg, 2.94 mmol) and Fmoc-OSu (1.04 g,3.08 mmol) in acetone (5 ml) was stirred at rt for 2 h. The solvent wasevaporated under reduced pressure. The residue was dissolved in EtOAc(50 ml) and washed with 5% NaHCO₃ (10 ml) and brine (10 ml). Organiclayer was dried (Na₂SO₄) and evaporated under reduced pressure. Theresidue was subjected to flash chromatography on CombiFlash® Companionunit equipped with RediSep® flash column (normal phase, 35-60 micronaverage particle size silicagel, 40 g, Teledyne Isco); flow rate=35ml/min; injection volume 2 ml (EtOAc); mobile phase A: hexane; mobilephase B: EtOAc; gradient 0-45% B in 1 h. Fractions containing thedesired product were combined and concentrated in vacuum to providecompound (14-7) (1.10 g, 2.4 mmol, 82% yield, mw 454.5) as colorlessoil. The oil solidified while under vacuum. (Alternatively, THF may beused as the solvent instead of acetone. It is also recommended to use 1eq. of DIEA to accelerate the reaction. Add the DIEA to the reactionafter the Fmoc reagent. The reaction can be concentrated, dissolved inEtOAc, washed with 1 M citrate or 5% NaHSO₄, water, 5% NaHCO₃, water,brine, dried (Na₂SO₄) and evaporated under reduced pressure. A standardsilica column purification can be accomplished eluting with 25% EtOAc inHexane to give 99% purity and 91% yield.)

(2S,3R)-2-Amino-3-Fmoc-aminobutyric acid methyl ester hydrochloride (14)

Compound (14-7) (1.10 g, 2.42 mmol) was dissolved in 4 N HCl/dioxane (8ml) at ambient temperature for 20 min. with stirring under nitrogen. Theresulting suspension was diluted with ether and the precipitate wasfiltered and washed with ether (3×10 mL). The compound (14) was dried invacuo to provide the hydrochloric salt (840 mg, 2.15 mmol, 89%, mw 354.4free base, HCl salt 390.4) as white solid. [M+H]=355.0. Retention time:4.11 min [Chromolith SpeedRod RP-18e C18 column (4.6×50 mm); flow rate1.5 ml/min; mobile phase A: 0.1% TFA/water; mobile phase B 0.1% TFA/ACN;gradient elution from 5% B to 100% B over 12 min, detection 254 nm]

Synthesis of Alternate Intermediate 009

(E)-methyl 4-(2-iodovinyl)benzoate (1)

Chromium chloride anhydrous (8.9 g, 73 mmol, 8 equiv) was dissolved in100 mL THF under nitrogen at 0° C. Triiodomethane (14.4 g, 37 mmol, 4equiv) in 150 mL THF was added dropwise in 10 min. Then compound (1S)(1.5 g, 9.1 mmol, 1 equiv) in 20 mL THF was added dropwise. The mixturewas stirred at 0° C. for 2 hours and then at room temperature for 1 morehour. The mixture was poured into iced water then extracted with EtOAc(100 mL×2). The organic phase was washed with 20% aq. Na₂S₂O₃ (100 mL),brine (100 mL) and dried (Na₂SO₄). The crude product was purified bychromatography on silica gel eluting with EtOAc/petroleum ether (0-2%)to give the title compound (1) (2.2 g, 85%). MS (m/z): [M+H]⁺=289.7. ¹HNMR (300 MHz, d₆-DMSO): 3.92 (s, 3H), 7.01˜7.05 (d, 1H), 7.33=7.34 (d,1H), 7.35˜7.36 (d, 1H), 7.44˜7.49 (d. 1H), 7.97˜7.98 (d, 1H), 7.99=8.00(d, 1H).

(E)-methyl 4-(4-(4-(morpholinomethyl)phenyl)but-1-en-3-ynyl)benzoate (2)

Compound (1) (2 g, 6.9 mmol, 1 equiv), diisopropylamine (0.7 g, 6.9mmol, 1 equiv) and PdCl₂(PPh₃)₂ (0.24 g, 0.34 mmol, 0.05 equiv) wasdissolved in 20 mL triethylamine at room temperature under nitrogen. Themixture was stirred for 15 min followed by addition of compound (004)(1.4 g, 6.9 mmol, 1 equiv). The mixture was stirred at room temperaturefor 12 hours. The reaction mixture was diluted with 50 mL EtOAc,filtered and concentrated. Then, the residue was diluted with 100 mlEtOAc and washed with water (100 mL) and brine (100 mL), dried (Na₂SO₄)and concentrated under reduced pressure. The crude product was purifiedby chromatography on silica gel eluting with EtOAc/petroleum ether(0-10%) to give the title compound (2) (1.5 g, 60%). MS (m/z):[M+H]⁺=362. ¹H NMR (300 MHz, d₆- DMSO): 2.3˜22.34 (d, 4H), 3.46 (s, 1H),3.5˜43.57 (t, 4H), 3.83 (s, 3H), 6.77=6.82 (d. 1H), 7.13=7.18 (d, 1H),7.32=7.34 (d, 2H), 7.4˜37.45 (d, 2H), 7.68˜7.71 (d, 2H), 7.90=7.93 (d,2H).

(E)-4-(4-(4-(morpholinomethyl)phenyl)but-1-en-3-ynyl)benzoic acid (009)

Compound (2) (1.5 g, 4.2 mmol, 1 equiv) was dissolved in a solvent of(THF:MeOH:H₂O=1:1:1, 20 mL) at room temperature. Then, lithium hydroxide(0.35 g, 6.4 mmol, 2 equiv) was added. The mixture was stirred for 12hours at room temperature. The pH was adjusted to 7 by acetic acid. Themixture was stirred for 1 hour. The solid was filtered and washed withwater (20 mL), ether 20 mL and dried in vacuo to give the target product(009) (0.9 g, 62%). MS (m/z): [M+H]⁺=348. ¹H NMR (300 MHz, d₆-DMSO):2.3˜22.35 (t, 4H), 3.47 (s, 2H), 3.54˜3.57 (t, 4H), 6.75˜6.80 (d. 1H),7.1˜27.18 (d, 1H), 7.3˜27.35 (d, 2H), 7.43=7.45 (d, 2H), 7.66=7.68 (d,2H), 7.88˜7.91 (d, 2H).

Synthesis of Alternate Intermediate (011)

(4-Iodophenyl)methanol (2)

To a solution of compound (1) (50 g, 0.21 mol, 1.0 equiv) in methanol(500 mL) was added with NaBH₄ (20.5 g, 0.5 mol, 2.5 equiv) slowly under10° C. for 1 h and stirred for 2 h. The reaction mixture wasconcentrated under reduced pressure, quenched with water (300 mL), andextracted with EtOAc (300 mL×3). The organic layer was dried overNa₂SO₄, filtered and concentrated under reduced pressure. The residuewas dried in vacuo overnight to provide the title compound (2) (49 g,97%).

Tert-butyl(4-iodobenzyloxy)dimethylsilane (3)

To a solution of compound (2) (9 g, 38.5 mmol, 1.0 equiv), TBSCl (5.8 g,38.5 mmol, 1.0 equiv) and imidazole (5.3 g, 77 mmol, 2.0 equiv) in DMF(18 mL) was stirred for 2 h at 45° C. The reaction mixture was dilutedwith CH₂Cl₂ (200 mL) and washed with water (200 mL×3). The organic layerwas dried over Na₂SO₄, filtered and concentrated under reduced pressure.The residue was dried under high vacuum overnight to produce the titlecompound (3) (11.6 g, 87%). ¹H NMR (300 MHz, d₆-DMSO): 0.05 (s, 6H),0.88 (s, 9H), 4.64 (s, 2H), 7.09˜7.11 (d. 2H), 7.66˜7.69 (d, 2H).

Tert-butyldimethyl(4-((trimethylsilyl)ethynyl)benzyloxy)silane (4)

Under nitrogen, ethynyltrimethylsilane (3.929 g, 37 mmol, 1.1 equiv) wasadded to a solution of compound (3) (11.6 g, 33.4 mmol 1.0 equiv),PdCl₂(PPh₃)₂ (2.345 g, 3.34 mmol, 0.1 equiv), CuI (0.636 g, 3.34 mmol,0.1 equiv) and i-Pr₂NH (10.05 mL) in THF (anhydrous, 200 ml) at roomtemperature. The mixture was allowed to reaction at ambient temperaturefor 3 h. Then, the precipitate was isolated. The filter cake was washedwith EtOAc (50 mL). The filtrate was concentrated under reducedpressure. The residue was dissolved in EtOAc (300 mL) and washed withwater (50 ml), 1M HCl aq. (50 ml), water (50 ml×2) and brine (50 ml).The organic layer was separated, dried (Na₂SO₄) and the filtrate wasconcentrated under reduced pressure. The residue was purified bychromatography (EtOAc/petroleum ether 0-2%) on silica gel to give thetitle compound (4) (9.2 g, 87%). ¹H NMR (300 MHz, d₆-DMSO): 0.10 (s,6H), 0.26 (s, 9H), 0.93 (s, 9H), 4.73 (s, 2H), 7.23˜7.26 (d. 2H),7.42˜7.44 (d, 2H).

Tert-butyl(4-ethynylbenzyloxy)dimethylsilane (5)

To a solution of compound (4) (5.2 g, 16.4 mmol, 1.0 equiv) in methanol(100 mL) was treated with KOH/methanol (0.457 g/5 mL) slowly below 10°C., then the mixture was allowed to react at ambient temperature for 5min. The reaction mixture was neutralized with acetic acid andconcentrated under reduced pressure. The residue was dissolved in EtOAc(100 mL). The organic layer was washed with 5% aq. Na₂CO₃ (50 ml×2),dried over Na₂SO₄, filtered and concentrated under reduced pressure. Theresidue was dried in vacuo overnight to provide the title compound (5)(4 g, 99%). ¹H NMR (300 MHz, d₆-DMSO): 0.04 (s, 6H), 0.87 (s, 9H), 4.09(s, 1H), 4.68 (s, 2H), 7.26˜7.29 (d. 2H), 7.4˜17.43 (d, 2H).

(E)-methyl4-(4-(4-((tert-butyldimethylsilyloxy)methyl)phenyl)but-1-en-3-ynyl)benzoate (010)

Under nitrogen, (E)-methyl 4-(2-iodovinyl)benzoate (1 g, 3.47 mmol, 1.0equiv) was added to a solution of compound (5) (0.855 g, 3.47 mmol, 1.0equiv), PdCl₂(PPh₃)₂ (0.244 g, 0.347 mmol, 0.1 equiv), CuI (0.066 g,0.347 mmol, 0.1 equiv) and i-Pr₂NH (2 mL) in THF (anhydrous, 10 mL) atroom temperature. The mixture was allowed to reaction at ambienttemperature for 3 h. Then, the precipitate was filtered. The filter cakewas washed with EtOAc. The filtrate and wash liquid were combined andconcentrated under reduced pressure. The residue was dissolved in EtOAc(30 ml) and washed with water (10 ml), 1M HCl aq. (10 ml), water (10ml×2) and brine (10 ml). The organic layer was separated, dried(Na₂SO₄), filtered and concentrated under reduced pressure. The residuewas purified by chromatography (EtOAc/petroleum ether 0-3%) on silicagel to give the target compound (010) (9.2 g, 86.6%). ¹H NMR (300 MHz,d₆-CDCl₃): 0.10 (s, 6H), 0.94 (s, 9H), 3.91 (s, 3H), 4.74 (s, 2H),6.46˜6.52 (d. 1H), 7.01˜7.07 (d, 1H), 7.26˜7.31 (t, 2H), 7.43˜7.49 (m,4H), 7.99˜8.02 (t, 2H).

Example 31 Step 1

A solution of ethyl 4-iodo-benzoate (2.43 g, 8.81 mmol) and propargylalcohol (543.3 mg, 9.69 mmol) in THF (35 mL) was purged by nitrogen for30 min, followed by addition of PdCl₂(PPh₃)₂ (74 mg, 0.106 mmol) and CuI(40 mg, 0.212 mmol). The reaction mixture was cooled to 0° C. andtriethyl amine (1.34 g, 13.22 mmol) was added. The reaction mixture waswarmed to rt and stirred overnight under N₂. The reaction mixture wasfiltered through a plug of celite and the filtrate was concentrated. Theresidue was treated with water (200 mL) and extracted with EtOAc (200mL×2). The combined organic layers were washed with water (200 mL×2),brine (200 mL) and dried (NaSO₄). The crude product was purified bychromatography on silica gel eluting with EtOAc/Hexane (0-50%) to give1.53 g, 85% yield) of the product (20).

Dess-Martin reagent (4.78 g, 11.25 mmol) was added to a solution ofcompound (20) (1.53 g, 7.5 mmol) in CH₂Cl₂ (20 mL) at 0° C. The reactionmixture was warmed to rt and stirred overnight. The reaction mixture wasdiluted with CH₂Cl₂ (200 mL), basified with NaHCO₃ (saturated) to pH=7and then washed with water (200 mL) and brine (200 mL), dried (NaSO₄)and concentrated under reduced pressure. The crude product was purifiedby chromatography on silica gel eluting with EtOAc/Hexane (0-40%) togive the desired product (21) (1.24 g, 81.8%).

Step 2

p-Toluenesulfonic acid (721 mg, 12% HOAc solution) was added to abenzene (30 ml) solution of 4-bromomethyl-benzaldhyde (1 g, 5.03 mmol)and ethylene glycol (623 mg, 10.06 mmol). The reaction was heated underrefluxing overnight. After cooled to room temp, the reaction mixture wasconcentrated under reduced pressure. The crude product was purified bychromatography on silica gel eluting with EtOAc/Hexane (0-20%) to givethe desired product (22) (1.13 g, 92.8%).

Triphenylphosphine (1.34 g, 5.11 mmol) was added to a 1,2-dichloroethane(10 ml) solution of compound (22) (1.13 g, 4.65 mmol). The reaction washeated to 70° C. for 4 hours. After cooling to room temp., the organicsolution was concentrated with rotavap. Diethyl ether (50 ml) was addedto the residue with vigorous stirring to precipitated the wittig salt.After filtration, the collected solid was washed with ether (3×50 ml)and dried under high vacuum to give compound (23) (2 g, 85% yield).

Step 3

To a solution of compound (23) (1.36 g, 2.69 mmol) in DCM (10 ml) wasadded DIPEA (694 mg, 5.38 mmol). The reaction mixture was stirred at rtfor 10 min., followed by addition of compound (21) (544 mg, 2.69 mmol)in DCM (10 ml) solution.

The resulting mixture was stirred at rt overnight and concentrated underreduced pressure. The residue was treated with EtOAc (100 ml), washedwith water (2×100 ml) and brine (100 ml), dried (NaSO₄) and concentratedunder reduced pressure. The crude product was purified by chromatographyon silica gel (0-20% EtOAc/Hexane) to give compound (24) (600 mg,63.9%).

Step 4

To a solution of compound (24) (718 mg, 2.35 mmol) in THF (6 ml) andHOAc (6 ml) was added NaBH₃CN (211 mg, 3.55 mmol). The reaction wasstirred at room temp. for 30 min. The reaction mixture was thenconcentrated under reduced pressure. The residue was diluted with EtOAc(200 ml) and treated with NaHCO₃ (sat'd) to pH=8. After separation, theaqueous layer was extracted with EtOAc (2×200 ml). The combined organiclayers was washed with H₂O (2×100 ml), brine (100 ml) and dried withNa₂SO₄. Concentrated by ratovap, the residue was purified bychromatography on silica gel (0-50% EtOAc/Hexane) to give compound (25a)(300 mg) and (25b) (187.5 mg) with 67% total yield.

Step 5

A DCM solution (5 ml) of compound 25 (300 mg, 0.98 mmol) and DIPEA (158mg, 1.23 mmol) was cooled to 0° C., followed by the addition MsCl (124mg, 1.08 mmol). The reaction was warmed to rt and stirred for 30 min.Morphline (171 mg, 1.96 mmol) was added to reaction and stirred for 2hours at rt. Diluted with EtOAc (200 ml), reaction mixture was washedwith H₂O (2×100 ml), brine (100 ml) and dried with Na₂SO₄. Concentratedunder reduced pressure, the crude product was purified by chromatographyon silica gel (0-50% EtOAc/Hexane) to give compound (27) (345 mg) in 94%yield.

Step 6

The desired product (28) (284 mg, 95%) was prepared with compound (26)(244 mg, 0.8 mmol), MsCl (80.5 mg, 0.88 mmol), morphline (106 mg, 1.22mmol) and DIPEA (100 mg, 0.88 mmol) according to the synthetic procedurefor the preparation of compound (27).

Step 7

To a mix solution (6 ml) of THF/MeOH/H₂O (1:1:1) of compound (27) (345mg, 0.92 mmol) was added LiOH (44 mg, 1.84 mmol). Reaction was stirredover at rt. HOAc was added to the reaction to pH=7. The reaction wasconcentrated under reduced pressure. The residue was re-dissolved inCH₃CN (1 ml) and H₂O (2 ml) and dried with lyophlizer to give crudeproduct (29) (482 mg, 66% pure, 100% yield).

Step 8

The desired product (30) (374 mg, 76% pure, 100% yield) was preparedwith compound (28) (284 mg, 0.76 mmol), LiOH (36 mg, 1.52 mmol)according to the synthetic procedure for the preparation of compound(29).

Step 9

The desired product (31) (65 mg, 48%, MS⁺=463.1) was prepared by thereaction of compound (29) (100 mg, 0.29 mmol), L-Threonine methyl ester(88 mg, 0.44 mmol), HATU (176 mg, 0.46 mmol) and DIPEA (187 mg, 1.45mmol) in DMF (2 mL) according to the synthetic procedure for thepreparation of compound (7).

Step 10

The desired product (32) (100 mg, 63%, MS⁺=548.1) was prepared by thereaction of compound (29) (100 mg, 0.29 mmol), methyl 3-Boc-2,3-diaminopropanoade (112 mg, 0.44 mmol), HATU (176 mg, 0.46 mmol) andDIPEA (187 mg, 1.45 mmol) in DMF (12 mL) according to the syntheticprocedure for the preparation of compound (7).

Step 11

The desired product (33) (70 mg, 52%, MS⁺=463.1) was prepared by thereaction of compound (30) (100 mg, 0.29 mmol), L-Threonine methyl ester(88 mg, 0.44 mmol), HATU (176 mg, 0.46 mmol) and DIPEA (187 mg, 1.45mmol) in DMF (2 mL) according to the synthetic procedure for thepreparation of compound (7).

Step 12

The desired product (34) (90 mg, 56.7%, MS⁺=548.1) was prepared by thereaction of compound (30) (100 mg, 0.29 mmol), Methyl 3-Boc-2,3-diaminopropanoade (112 mg, 0.44 mmol), HATU (176 mg, 0.46 mmol) andDIPEA (187 mg, 1.45 mmol) in DMF (12 mL) according to the syntheticprocedure for the preparation of compound (7).

Step 13

The desired product (35) (20 mg, 14.9%, MS⁺=464.1) was prepared by thereaction of compound (31) (65 mg, 0.14 mmol) and NH₂OH.HCl (96.6 mg, 1.4mmol) and NaOMe (115 mg, 2.1 mmol) in MeOH (5 ml) according to thesynthetic procedure for the preparation of compound (8).

Step 14

The desired product (36) (30 mg, 30%, MS⁺=464.1) was prepared by thereaction of compound (32) (100 mg, 0.182 mmol) and NH₂OH.HCl (126 mg,1.8 mmol) and NaOMe (148.5 mg, 2.7 mmol) in MeOH (5 ml) according to thesynthetic procedure for the preparation of compound (8).

Step 15

The desired product (37a) (20 mg, 28.4%, MS⁺=464.1) was prepared by thereaction of compound (33) (70 mg, 0.152 mmol) and NH₂OH.HCl (104 mg,1.52 mmol) and NaOMe (125 mg, 2.28 mmol) in MeOH (5 ml) according to thesynthetic procedure for the preparation of compound (8).

Step 16

The desired product (37b) (40 mg, 44.5%, MS⁺=548.1) was prepared by thereaction of compound (34) (90 mg, 0.164 mmol) and NH₂OH.HCl (114 mg,1.65 mmol) and NaOMe (136.1 mg, 2.47 mmol) in MeOH (5 ml) according tothe synthetic procedure for the preparation of compound (8).

Step 17

To a solution of compound (36) (30 mg, 0.055 mmol) in CH₂Cl₂ (1 ml) wasadded HCl (1 ml, 4 N in dioxane). The reaction mixture was stirred atrt. for 2 hrs. and concentrated under reduced pressure. The solidresidue was purified by prep. HPLC to give compound (38) (10 mg, 40%yield, MS⁺=449.1) as double TFA salt.

Step 18

To a solution of compound (37b) (40 mg, 0.073 mmol) in CH₂Cl₂ (1 ml) wasadded HCl (1 ml, 4 N in dioxane). The reaction mixture was stirred atrt. for 2 hrs. and concentrated under reduced pressure. The solidresidue was purified by prep. HPLC to give compound (39) (15 mg, 45.8%yield, MS⁺=449.1) as double TFA salt.

Step 19

The desired products (40) and (41) may be prepared according to theforegoing synthetic procedures.

Example 32

Step 1

NaBH(OAc)₃ (2.28 g, 0.76 mmol) was added at 0° C. to a solution of4-ethylnyl-benzylaldehyde (1.0 g, 7.69 mmol) and morpholine (462 mg,5.37 mmol) in THF (20 mL). The reaction mixture was then warmed to rtand stirred overnight. The solvent was removed (RV) and the residue wasextracted with EtOAc (2×) from aqueous NaHCO₃ (pH=8˜9). The combinedorganic extracts were dried (Na₂SO₄) and concentrated to dryness (RV).Column chromatography (silica gel, EtOAc/DCM 0-50%) yielded compound(40) (1.31 g, 84.8% yield, M+H⁺=202.0).

Step 2

A solution of methyl 4-formylbenzoate (1 g, 6.09 mmol) and p-TSA (12% inAcOH, 104 mg, 0.609 mmol) in benzene solution (50 mL) was heated atreflux for 3 hours. The reaction mixture was cooled to rt, diluted withEtOAc (200 ml) and washed with water (200 mL×2) and brine (200 mL). Thecrude product was purified by chromatography on silica gel eluting withEtOAc/Hexane (0-50%) to give compound (41) (1.1 g, 87%)

Step 3

NaHMDS was added to a solution of compound (40) in THF at 0° C. Thereaction was stirred for 1 hour, followed by addition of compound (41)in THF at 0° C. The reaction mixture was warmed to rt and stirredovernight. The reaction mixture was treated with NH₄Cl (saturated) andextracted with EtOAc. The combined organic layers were washed with brine(20 ml), dried (NaSO₄) and concentrated under reduced pressure. Thecrude product was purified by chromatography on silica gel.

Further Steps

Example 33

Example 34

Example 35

Example 36

The following compounds may be synthesized as described in this, and theforegoing, Example.

Examples 37A and 37B

Example 38

Example 39

The following compound may be synthesized as described in this Example.

Example 40

Example 41N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-[1-(4-morpholin-4-ylmethyl-phenyl)-piperidin-4-yl]-benzamide

Synthesis of 4-[1-(4-formyl-phenyl)-piperidin-4-yl]-benzoic acid methylester (U

A mixture of 4-(4-methoxycarboxyphenyl) piperidine HCl (256 mg, 1.0mmol), 4-fluoro-benzaldehyde (105 μl, 1.0 mmol) and K₂CO₃ (250 mg, 1.8mmol) in DMF (5 ml) was stirred at ambient temperature overnight.Reaction was diluted with water (30 ml). Formed precipitate wasfiltrated, washed with water (20 ml) and ether (20 ml), and dried invacuum overnight to provide target compound (1) (203 mg, 63%) as whitesolid. LC-MS [M+H] 324.3 (C₂₀H₂₁NO₃+H, requires 324.41).

Synthesis of4-[1-(4-morpholin-4-ylmethyl-phenyl)-piperidin-4-yl]-benzoic acid methylester (2)

A mixture of compound (1) (200 mg, 0.62 mmol), NaBH(OAc)₃ (158 mg, 0.75mmol) and morpholine (60 μl, 0.68 mmol) in DCM (5 ml) was stirred atambient temperature overnight. Reaction was quenched with 5% aq. NaHCO₃(15 ml) and extracted with EtOAc (30 ml×2). Organic layer was washedwith brine (20 ml) and dried over MgSO₄ (anh.). Solvent was evaporatedin vacuum. Residue was dried in vacuum overnight to provide targetcompound (2) (225 mg, 92%) as yellowish solid. LC-MS [M+H] 308.1(C₂₄H₃₀N₂O₃+H, requires 395.53).

Synthesis of4-[1-(4-morpholin-4-ylmethyl-phenyl)-piperidin-4-yl]-benzoic acid (3)

A solution of compound (2) (225 mg, 0.57 mmol) and 1 M aq. NaOH (10 ml,10 mmol) in dioxane (6 ml) was stirred at ambient temperature overnight.Reaction mixture was acidified with 1 M aq. HCl to pH˜6. Volatilesolvent was evaporated in vacuum. Formed precipitate was filtrated andwashed with water (10 ml) and cold ether (5 ml) and dried in vacuumovernight to provide di-hydrochloric salt of target material (3) (211mg, 89%) as off-white solid. LC-MS [M+H] 381.4 (C₂₃H₂₈N₂O₃+H, requires381.50).

Synthesis of(2S,3R)-3-hydroxy-2-{4-[1-(4-morpholin-4-ylmethyl-phenyl)-piperidin-4-yl]-benzoylamino}-butyricacid methyl ester (4)

A solution of compound (3) (70 mg, 0.15 mmol), HATU (63 g, 0.17 mmol)and DIEA (131 μl, 0.75 mmol) in DMF (800 μl) was maintained at ambienttemperature for 10 min followed by the addition of H-Thr-OMehydrochloride (37 mg, 0.22 mmol). Reaction mixture was stirred atambient temperature overnight followed by the dilution with EtOAc (100ml). Solution was extracted with water (20 ml×2) and brine (20 ml).Organic layer was dried over MgSO₄ and evaporated. Residue was dried invacuum to provide target compound (4) (73 mg, 99%) as yellow solid.LC-MS [M+H] 496.3 (C₂₈H₃₇N₃O₅+H, requires 496.63).

Synthesis ofN-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-[1-(4-morpholin-4-ylmethyl-phenyl)-piperidin-4-yl]-benzamide(41-5)

A solution of hydroxylamine hydrochloride (63 mg, 0.9 mmol) in MeOH (2ml) was cooled to −5° C. followed by the addition of 25% NaOMe/MeOH (308μl, 4.5 mmol) under nitrogen. Reaction mixture was stirred at −5° C. foradditional 5 min, cooled to −20° C., and solution of compound 4 (73 mg,0.15 mmol) in THF/MeOH (1:1, 2 ml) was added dropwise over the period of5 min. Temperature of reaction mixture was raised to ambient. Completionof the reaction was monitored by LC-MS. Reaction mixture was acidifiedwith 1 N aq. HCl to pH˜7 and evaporated in vacuum. Residue was dissolvedin DMSO (500 μl) and subjected to HPLC purification. [Phenomenex GeminiC-18 column, 110 Å (30×100 mm); flow rate=20 ml/min; mobile phase A:100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1% TFA; gradientelution from 8% B to 28% B in 60 min., detection 254 nm]. Fractionscontaining the desired product were combined and lyophilized to providedi-trifluoroacetic salt of target product (41-5) as white solid. LC-MS[M+H] 497.7 (C₂₇H₃₆N₄O₅+H, requires 497.62).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (41-5) 0.15 76 70 100 497.7 1.97 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm]

Example 42N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperidin-1-yl]-benzamide

Synthesis of 4-(4-hydroxymethyl)-piperidine-1-carboxylicacid tert-butylester (1)

A solution of N-Boc-4-(4-carboxyphenyl) piperidine (305 mg, 1.0 mmol)and 1 M BH₃ THF (3.1 ml, 3.1 mmol) in THF (3 ml) was maintained undernitrogen at ambient temperature overnight. Reaction was quenched with 5%aq. NH₄Cl (5 ml) and extracted with EtOAc (30 ml×2). Organic layer waswashed with 5% NaHCO₃ (20 ml) and brine (20 ml) and dried over MgSO₄.Solvent was evaporated in vacuum. Residue was dried in vacuum overnightto provide target compound (1) (272 mg, 93%) as white solid. LC-MS [M+H]292.3 (C₁₇H₂₅NO₃+H, requires 292.40).

Synthesis of 4(4-Morpholin-4-ylmethyl-phenyl)-piperidine-1-carboxylicacid tert-butyl ester (2)

A solution of compound (1) (272 mg, 0.93 mmol) and DIEA (198 μl, 1.14mmol) in DCM (5 ml) was cooled to 0° C. followed by the addition of MsCl(87 μl, 1.12 mmol). Reaction mixture was maintained at 0° C. for 10 minfollowed by the addition of mixture of DIEA (179 μl, 1.1 mmol) andmorpholine (90 μl, 1.1 mmol). Temperature of the reaction mixture wasallowed to rise to ambient. Reaction mixture was maintained at ambienttemperature overnight and diluted with EtOAc (80 ml). Organic layer waswashed with 5% NaHCO₃ (20 ml) and brine (20 ml) and dried over MgSO₄(anh.). Solvent was evaporated in vacuum. Residue was dried in vacuumovernight to provide target compound (2) (315 mg, 94%) as brownish oil.LC-MS [M+H] 361.4 (C₂₁H₃₂N₂O₃+H, requires 361.51).

Synthesis of 4-(4-piperidin-4-yl-benzyl)-morpholine (3)

A solution of compound (2) (315 mg, 1.13 mmol), TFA (3 ml, 40 mmol) inDCM (3 ml) was maintained at ambient temperature for 3 h. Solvents wereevaporated in vacuum. Residue was dissolved in DCM (1 ml) and 1 MHCl/ether (50 ml) was added. Formed precipitate was filtrated, washedwith ether and dried in vacuum overnight to provide di-hydrochloric saltof target compound (3) (281 mg, 91%) as off-white solid. LC-MS [M+H]261.1 (C₁₆H₂₄N₂O+H, requires 261.39).

Synthesis of4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperidin-1-yl]-benzoic acid methylester (4)

A mixture of compound (3) (261 mg, 0.78 mmol), 4-methoxycarbonylphenyl)boronic acid (388 mg, 2.16 mmol), Cu(OAc)₂ (216 mg, 1.19 mmol),molecular sieves (4 Å, 200 mg) and Et₃N (750 μl, 5.6 mmol) in DCE (5 ml)was stirred at ambient temperature overnight under nitrogen. Formedprecipitate was filtrated. Filtrate was diluted with EtOAc (100 ml) andextracted with water (20 ml×2) and brine (20 ml). Solvent was evaporatedin vacuum. Residue was dissolved in DMSO (1 ml) and subjected to HPLCpurification. [YMC-Pack ODS-A C-18 column (30×100 mm); flow rate=10ml/min; injection volume 1.0 ml; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 10% B to 60% Bin 70 min., detection 254 nm]. Fractions containing the desired productwere combined and concentrated in vacuum. The residue was dissolved inEtOAc (25 ml), extracted with 5% aq. NaHCO₃ (20 ml) and brine (20 ml)and dried over MgSO₄ (anh.). Solvent was evaporated in vacuum. Residuewas dried in vacuum overnight to provide target product (4) (24 mg, 6%)as white solid. LC-MS [M+H] 394.8 (C₂₄H₃₀N₂O₃+H, requires 395.53).

Synthesis of4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperidin-1-yl]-benzoic acid (5)

A solution of compound (4) (24 mg, 0.06 mmol) and 1 M aq. NaOH (500 μl,0.5 mmol) in dioxane (500 μl) was stirred at ambient temperatureovernight. Reaction mixture was acidified with 1 M aq. HCl to pH˜2.Formed precipitate was filtrated and washed with water (10 ml) and driedin vacuum overnight to provide di-hydrochloric salt of target material(5) (24 mg, 88%) as off-white solid. LC-MS [M+H] 381.4 (C₂₃H₂₈N₂O₃+H,requires 381.50).

Synthesis of(2S,3R)-3-hydroxy-2-{4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperidin-1-yl]-benzoylamino}-butyricacid methyl ester (6)

A solution of compound (5) (24 mg, 0.053 mmol), HATU (25 g, 0.066 mmol)and DIEA (52 μl, 0.3 mmol) in DMF (600 μl) was maintained at ambienttemperature for 10 min followed by the addition of H-Thr-OMehydrochloride (14 mg, 0.08 mmol). Reaction mixture was stirred atambient temperature overnight followed by the dilution with EtOAc (100ml). Solution was extracted with water (20 ml×2) and brine (20 ml).Organic layer was dried over MgSO₄ and evaporated. Residue was dried invacuum to provide target compound (6) (26 mg, 99%) as yellow solid.LC-MS [M+H] 496.4 (C₂₈H₃₇H₃O₅+H, requires 496.63).

Synthesis ofN-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperidin-1-yl]-benzamide(42-7)

A solution of hydroxylamine hydrochloride (24 mg, 0.34 mmol) in MeOH (2ml) was cooled to −5° C. followed by the addition of 25% NaOMe/MeOH (117μl, 0.51 mmol) under nitrogen. Reaction mixture was stirred at −5° C.for additional 5 min, cooled to −20° C., and solution of compound (6)(26 mg, 0.053 mmol) in THF/MeOH (1:1, 2 ml) was added dropwise over theperiod of 5 min. Temperature of reaction mixture was raised to ambient.Completion of the reaction was monitored by LC-MS. Reaction mixture wasacidified with 1 N aq. HCl to pH˜7 and evaporated in vacuum. Residue wasdissolved in DMSO (500 μl) and subjected to HPLC purification.[Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flow rate=20 ml/min;mobile phase A: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1%TFA; gradient elution from 8% B to 28% B in 60 min., detection 254 nm].Fractions containing the desired product were combined and lyophilizedto provide di-trifluoroacetic salt of target product 42-7 as whitesolid. LC-MS [M+H] 497.5 (C₂₇H₃₆N₄O₅+H, requires 497.62).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (42-7) 0.053 17.5 46 99.8 497.5 2.22 *[ChromolithSpeedRod RP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobilephase A: 0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elutionfrom 5% B to 100% B over 9.6 mm, detection 254 nm]

Example 43N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperazin-1-yl]-benzamide

Synthesis of 4-(4-formyl-phenyl)-piperazine-1-carboxylic acid tert-butylester (1)

A mixture of 4-(tert-butyl-1-piperazinecarboxylate (556 mg, 3.0 mmol),4-fluoro-benzaldehyde (315 μl, 3.0 mmol) and K₂CO₃ (514 mg, 3.7 mmol) inDMF (5 ml) was stirred at ambient temperature overnight. Reaction wasdiluted with water (30 ml) and extracted with EtOAc (50 ml×2). Organiclayer was washed with water (20 ml), 1 M aq. HCl (20 ml), water (20ml×2) and brine (20 ml) and dried over MgSO₄ (anh.). Solvent wasevaporated in vacuum. Residue was triturated with hexane. Formedprecipitate was filtrated and dried in vacuum overnight to providetarget compound (1) (342 mg, 39%) as off-white solid. LC-MS [M+H] 291.2(C₁₆H₂₂N₂O₃+H, requires 291.38).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenyl)-piperazine-1-carboxylicacid tert-butyl ester (2)

A mixture of compound (1) (342 mg, 1.18 mmol), NaBH(OAc)₃ (300 mg, 1.42mmol) and morpholine (113 μl, 1.3 mmol) in DCM (5 ml) was stirred atambient temperature overnight. Reaction was quenched with 5% aq. NaHCO₃(15 ml) and extracted with EtOAc (30 ml×2). Organic layer was washedwith brine (20 ml) and dried over MgSO₄ (anh.). Solvent was evaporatedin vacuum. Residue was dried in vacuum overnight to provide targetcompound (2) (408 mg, 96%) as white solid. LC-MS [M+H] 361.9(C₂₀H₃₁N₃O₃+H, requires 362.50).

Synthesis of 4-(4-piperazin-1-yl-benzyl)-morpholine (3)

A solution of compound (2) (408 mg, 1.13 mmol), TFA (5 ml, 68 mmol) inDCM (5 ml) was maintained at ambient temperature for 40 min. Solventswere evaporated in vacuum. Residue was dissolved in 1 M aq. NaOH (10 ml)and extracted with EtOAc (30 ml×2). Organic layer was washed with brine(20 ml) and dried over MgSO₄ (anh.). Solvent was evaporated in vacuum.Residue was dried in vacuum overnight to provide target compound (3)(212 mg, 72%) as yellowish solid. LC-MS [M+H] 262.3 (C₁₅H₂₃N₃O+H,requires 262.38).

Synthesis of4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperazin-1-yl]-benzoic acid methylester (4)

A mixture of compound (3) (140 mg, 0.54 mmol), methyl 4-fluorobenzoate(76 μl, 0.59 mmol) and DIEA (141 μl 1, 0.81 mmol) in DMSO (2 ml) wasirradiated in microwave oven (max. power 250 W, 160° C.) for 2 h andcooled to ambient temperature. Reaction mixture was diluted with water(30 ml). Formed precipitate was filtrated and dissolved in EtOAc (100ml). Solution was washed with 5% aq. NaHCO₃ (20 ml) and brine (20 ml)and dried over MgSO₄ (anh.). Solvent was evaporated in vacuum. Residuewas dried in vacuum overnight to provide target compound (4) (98 mg,46%) as yellowish solid. LC-MS [M+H] 396.2 (C₂₃H₂₉N₃O₃+H, requires396.52).

Synthesis of4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperazin-1-yl]-benzoic acid (5)

A solution of compound (4) (98 mg, 0.57 mmol) and 1 M aq. NaOH (2 ml, 2mmol) in dioxane (2 ml) was stirred at ambient temperature overnight.Reaction mixture was acidified with 1 M aq. HCl to pH˜6 and evaporatedin vacuum. Residue was dissolved in DMSO (1 ml) and subjected to HPLCpurification. [YMC-Pack ODS-A C-18 column (30×100 mm); flow rate=10ml/min; injection volume 1.0 ml; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 0% B to 45% Bin 72 min., detection 254 nm]. Fractions containing the desired productwere combined and concentrated in vacuum. The residue was dissolved ini-PrOH (15 ml) and evaporated in vacuum. Obtained residue was dissolvedin i-PrOH (5 ml) and 1 M HO/ether (50 ml) was added. Precipitate wasfiltrated and dried in vacuum to provide tri-hydrochloric salt of targetproduct (5) (30 mg, 24%) as off-white solid. LC-MS [M+H] 382.6(C₂₂H₂₇N₃O₃+H, requires 382.49).

Synthesis of(2S,3R)-3-hydroxy-2-{4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperazin-1-yl]-benzoylamino}-butyricacid methyl ester (6)

A solution of compound (5) (30 mg, 0.06 mmol), HATU (25 g, 0.066 mmol)and DIEA (63 μl, 0.36 mmol) in DMF (500 μl) was maintained at ambienttemperature for 10 min followed by the addition of H-Thr-OMehydrochloride (14 mg, 0.084 mmol). Reaction mixture was stirred atambient temperature overnight followed by the dilution with EtOAc (100ml). Solution was extracted with water (20 ml×2) and brine (20 ml).Organic layer was dried over MgSO₄ and evaporated. Residue was dried invacuum to provide target compound (6) (30 mg, 99%) as yellow solid.LC-MS [M+H] 479.0 (C₂₇H₃₆N₄O₅+H, requires 497.62).

Synthesis ofN-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-[4-(4-morpholin-4-ylmethyl-phenyl)-piperazin-1-yl]-benzamide(43-7)

A solution of hydroxylamine hydrochloride (63 mg, 0.36 mmol) in MeOH (2ml) was cooled to −5° C. followed by the addition of 25% NaOMe/MeOH (123μl, 0.36 mmol) under nitrogen. Reaction mixture was stirred at −5° C.for additional 5 min, cooled to −20° C., and solution of compound (6)(30 mg, 0.06 mmol) in THF/MeOH (1:1, 2 ml) was added dropwise over theperiod of 5 min. Temperature of reaction mixture was raised to ambient.Completion of the reaction was monitored by LC-MS. Reaction mixture wasacidified with 1 N aq. HCl to pH˜7 and evaporated in vacuum. Residue wasdissolved in DMSO (500 μl) and subjected to HPLC purification.[Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flow rate=20 ml/min;mobile phase A: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1%TFA; gradient elution from 8% B to 28% B in 60 min., detection 254 nm].Fractions containing the desired product were combined and lyophilizedto provide tri-trifluoroacetic salt of target product (43-7) as whitesolid. LC-MS [M+H] 498.3 (C₂₆H₃₅N₅O₅+H, requires 498.61).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (43-7) 0.06 15.3 31 100 498.61 2.64 *[Chromolith SpeedRodRP-18e C18 column (4.6 x 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm]The following compounds may be synthesized as described in this Example.

Example 44

Example 45

Example 46N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-benzylsulfanyl)-benzamide

Synthesis of 4-(4-morpholin-4-ylmethyl-benzylsulfanyl)-benzoic acidmethyl ester (1)

A solution of (4-(morpholinomethyl)phenyl)methanol (207 mg, 1.0 mmol)and DIEA (192 μl, 1.1 mmol) in DCM (3 ml) was cooled to 0° C. followedby the addition of MsCl (94 μl, 1.2 mmol). Reaction mixture wasmaintained at 0° C. for 1 h followed by the addition of mixture of DIEA(192 μl, 1.1 mmol) and methyl-4-mercaptobenzoate (185 mg, 1.1 mmol).Temperature of the reaction mixture was allowed to rise to ambient.Reaction mixture was maintained at ambient temperature 40 min anddiluted with EtOAc (80 ml). Organic layer was washed with 5% NaHCO₃ (20ml) and brine (20 ml) and dried over MgSO₄ (anh.). Solvent wasevaporated in vacuum. Residue (1) was used as is for the nexttransformation. LC-MS [M+H] 358.1 (C₂₀H₂₃NO₃S+H, requires 358.49).

Synthesis of 4-(4-morpholin-4-ylmethyl-benzylsulfanyl)-benzoic acid (2)

To a solution of compound (1) (1.0 mmol) in dioxane (1 ml) 1 M aq. NaOH(1.5 ml, 1.5 mmol) was added. Reaction mixture was maintained at ambienttemperature overnight and acidified with 1 M aq. HCl to pH˜5. Solventswere evaporated. Residue was dissolved in DMSO (1 ml) and subjected toHPLC purification. [YMC-Pack ODS-A C-18 column (30×100 mm); flow rate=10ml/min; injection volume 1.5 ml; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 5% B to 50% Bin 70 min., detection 254 nm]. Fractions containing the desired productwere combined and concentrated in vacuum. The residue was dissolved ini-PrOH (15 ml) and evaporated in vacuum. Obtained residue was dissolvedin i-PrOH (5 ml) and 1 M HCl/ether (50 ml) was added. Precipitate wasfiltrated and dried in vacuum to provide hydrochloric salt of targetproduct (2) (161 mg, 54%) as white solid. LC-MS [M+H] 344.1(C₁₉H₂₁NO₃S+H, requires 344.46).

Synthesis of(2S,3R)-3-hydroxy-2-[4-(4-morpholin-4-ylmethyl-benzylsulfanyl)-benzoylamino]-butyric acid methyl ester (3)

A solution of compound (2) (34 mg, 0.09 mmol), HATU (34 g, 0.09 mmol)and DIEA (63 μl, 0.36 mmol) in DMF (800 μl) was maintained at ambienttemperature for 10 min followed by the addition of H-Thr-OMehydrochloride (18 mg, 0.11 mmol). Reaction mixture was stirred atambient temperature overnight followed by the dilution with EtOAc (100ml). Solution was extracted with water (20 ml×2) and brine (20 ml).Organic layer was dried over MgSO₄ and evaporated. Residue was dried invacuum to provide target compound (3) (40 mg, 98%) as yellow solid.LC-MS [M+H] 459.2 (C₂₄H₃₀N₂O₅S+H, requires 459.59).

Synthesis ofN-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-benzylsulfanyl)-benzamide(46-4)

A solution of hydroxylamine hydrochloride (19 mg, 0.27 mmol) in MeOH (2ml) was cooled to −5° C. followed by the addition of 25% NaOMe/MeOH (94μl, 0.41 mmol) under nitrogen. Reaction mixture was stirred at −5° C.for additional 5 min, cooled to −20° C., and solution of compound (3)(40 mg, 0.09 mmol) in THF/MeOH (1:1, 2 ml) was added dropwise over theperiod of 5 min. Temperature of reaction mixture was raised to ambient.Completion of the reaction was monitored by LC-MS. Reaction mixture wasacidified with 1 N aq. HCl to pH˜7 and evaporated in vacuum. Residue wasdissolved in DMSO (500 W) and subjected to HPLC purification.[Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flow rate=20 ml/min;mobile phase A: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1%TFA; gradient elution from 8% B to 28% B in 60 min., detection 254 nm].Fractions containing the desired product were combined and lyophylisedto provide trifluoroacetic salt of target product (46-4) (25.8 mg, 50%)as white solid. LC-MS [M+H] 460.2 (C₂₃H₂₉N₃O₅S+H, requires 460.58).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (46-4) 0.09 25.8 50 98.7 460.2 3.11 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 mm, detection 254 nm]

Compounds (5) and (6)

Compounds (5) and (6) may be synthesized from compound 4 using OXONEoxidation in dichloromethane.

Example 47N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylsulfanyl)-benzamide-4-ylmethyl-phenylethynyl)-benzamide

Synthesis of 4-(4-iodo-benzyl)-morpholine (1)

To a solution of morpholine (353 μl, 4.06 mmol) in DMSO (5 ml) was added4-iodobenzylbromide (402 mg, 1.35 mmol). Reaction mixture was stirred atambient temperature overnight. Reaction mixture was diluted with 5%NaHCO₃ (100 ml) and extracted with EtOAc (50 ml×2). Organic layer waswashed with brine (30 ml), dried over MgSO₄ (anh.) and evaporated invacuum. Residue was dried in vacuum overnight at ambient temperature toprovide target material (1) (344 g, 84%) as white solid. LC-MS [M+H]304.1 (C₁₁H₁₄INO+H, requires 304.15).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenylsulfanyl)-benzoic acidmethyl ester (2)

A mixture of compound (1) (260 mg, 0.86 mmol), methyl-4-mercaptobenzoate(159 mg, 0.94 mmol), CuI (16.3 mg, 0.086 mmol and K₂CO₃ (237 mg, 1.72mmol) in NMP (2 ml) was stirred was irradiated in microwave oven (max.power 250 W, 160° C.) for 40 min and cooled to ambient temperature.Reaction mixture was diluted with EtOAc (80 ml) and extracted with 5%aq. NaHCO₃ (20 ml) and brine (20 ml) and dried over MgSO₄ (anh.).Solvent was evaporated in vacuum. Residue was dried in vacuum overnightto provide target compound (2) (289 mg, 98%) as yellowish solid. LC-MS[M+H] 344.3 (C₁₉H₂₁NO₃S+H, requires 344.46).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenylsulfanyl)-benzoic acid (3)

To a solution of compound (2) (289 mg, 0.84 mmol) in dioxane (500 μl) 1M aq. NaOH (1 ml) was added. Reaction mixture was maintained at ambienttemperature for 1.5 h, acidified with 1 M aq. HCl to pH˜5 and evaporatedin vacuum. Residue was dissolved in DMSO (1 ml) and subjected to HPLCpurification. [YMC-Pack ODS-A C-18 column (30×100 mm); flow rate=10ml/min; injection volume 1.0 ml; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 5% B to 50% Bin 72 min., detection 254 nm]. Fractions containing the desired productwere combined and concentrated in vacuum. The residue was dissolved ini-PrOH (15 ml) and evaporated in vacuum. Obtained residue was dissolvedin i-PrOH (5 ml) and 1 M HCl/ether (50 ml) was added. Precipitate wasfiltrated and dried in vacuum to provide hydrochloric salt of targetproduct (3) (77 mg, 25%) as yellowish solid. LC-MS [M+H] 330.1(C₁₈H₁₉NO₃S+H, requires 330.43).

Synthesis of(2S,3R)-3-hydroxy-2-[4-(4-morpholin-4-ylmethyl-phenylsulfanyl)-benzoylamino]-butyric acid methyl ester (4)

A solution of compound (3) (33 mg, 0.09 mmol), HATU (34 g, 0.09 mmol)and DIEA (63 μl, 0.36 mmol) in DMF (800 μl) was maintained at ambienttemperature for 10 min followed by the addition of compound H-Thr-OMehydrochloride (18 mg, 0.11 mmol). Reaction mixture was stirred atambient temperature overnight followed by the dilution with EtOAc (80ml). Solution was extracted with water (20 ml×2) and brine (20 ml).Organic layer was dried over MgSO₄ and evaporated. Residue was dried invacuum to provide target compound (4) (39 mg, 98%) as brown solid. LC-MS[M+H] 445.3 (C₂₃H₂₈N₂O₅S+H, requires 445.56).

Synthesis ofN-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylsulfanyl)-benzamide(47-5)

A solution of hydroxylamine hydrochloride (39 mg, 0.54 mmol) in MeOH(anh, 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (188 μl, 0.81 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (4) (0.09 mmol) in THF/MeOH (1:1, 2 ml) was added dropwiseover the period of 5 min. Temperature of reaction mixture was raised toambient. Completion of the reaction was monitored by LC-MS. Aftercompletion reaction mixture was acidified with 1 N aq. HCl to pH˜7 andevaporated in vacuum. Residue was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophylised to provide trifluoroacetic salt of targetproduct (47-5) (25.8 mg, 52%) as white solid. LC-MS [M+H] 446.2(C₂₂H₂₇N₃O₅S+H, requires 446.55).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (47-5) 0.09 25.8 52 98.7 446.2 3.11 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm

Compound (6)

Compound (6) may be synthesized from compound (5) using OXONE oxidationin dichloromethane.

Example 48N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-benzyl)-benzamide

Synthesis of 4-(4-hydroxymethyl-benzyl)-benzoic acid methyl ester (1)

A mixture of 4-(hydroxymethyl)phenylboronic acid (304 mg, 2.0 mmol),methyl 4-(bromomethyl)-benzoate (229 mg, 1.0 mmol), Pd(PPh₃)₄ (12 mg,0.01 mmol) and 1 M aq. K₂CO₃ (400 μl) in ACN (800 μl) was irradiated inmicrowave oven (max. power 250 W, 160° C.) for 7 min and cooled toambient temperature. Reaction mixture was diluted with EtOAc (100 ml)and extracted with 5% aq. NaHCO₃ (30 ml) and brine (30 ml). Organiclayer was dried over MgSO₄ (anh.). Solvent was evaporated in vacuum.Residue was dried in vacuum overnight to provide target product (1) (224mg, 88%) as yellow oil.

Synthesis of 4-(4-morpholin-4-ylmethyl-benzyl)-benzoic acid methyl ester(2)

A solution of compound (1) (204 mg, 0.8 mmol) and DIEA (191 μl, 1.1mmol) in DCM (2 ml) was cooled to 0° C. followed by the addition of MsCl(86 μl, 1.1 mmol). Reaction mixture was maintained at 0° C. for 1 hfollowed by the addition of mixture of DIEA (191 μl, 1.1 mmol) andmorpholine (131 μl, 1.5 mmol). Temperature of the reaction mixture wasallowed to rise to ambient. Reaction mixture was maintained at ambienttemperature overnight and diluted with EtOAc (80 ml). Organic layer waswashed with 5% NaHCO₃ (20 ml) and brine (20 ml) and dried over MgSO₄(anh.). Solvent was evaporated in vacuum. Residue was dissolved in DMSO(1 ml) and subjected to HPLC purification. [YMC-Pack ODS-A C-18 column(30×100 mm); flow rate=15 ml/min; injection volume 1.0 ml; mobile phaseA: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1% TFA; gradientelution from 15% B to 70% B in 70 min., detection 254 nm]. Fractionscontaining the desired product were combined and concentrated in vacuum.The residue was dissolved in EtOAc (25 ml), extracted with 5% aq. NaHCO₃(20 ml) and brine (20 ml) and dried over MgSO₄ (anh.). Solvent wasevaporated in vacuum. Residue was dried in vacuum overnight to providetarget product (2) (73 mg, 28%) as white solid. LC-MS [M+H] 326.3(C₂₀H₂₃NO₃+H, requires 326.42).

Synthesis of 4-(4-morpholin-4-ylmethyl-benzyl)-benzoic acid (3)

To a solution of compound (2) (73 mg, 0.22 mmol) in dioxane (500 μl) 1 Maq. NaOH (1.5 ml, 1.5 mmol) was added. Reaction mixture was maintainedat ambient temperature overnight and acidified with 1 M aq. HCl to pH˜5.Solvents were evaporated. Residue was dissolved in DMSO (1 ml) andsubjected to HPLC purification. [YMC-Pack ODS-A C-18 column (30×100 mm);flow rate=15 ml/min; injection volume 1 ml; mobile phase A: 100% water,0.1% TFA; mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 5% Bto 50% B in 70 min., detection 254 nm]. Fractions containing the desiredproduct were combined and concentrated in vacuum. The residue wasdissolved in i-PrOH (15 ml) and evaporated in vacuum. Obtained residuewas dissolved in i-PrOH (5 ml) and 1 M HCl/ether (50 ml) was added.Precipitate was filtrated and dried in vacuum to provide hydrochloricsalt of target product (3) (38 mg, 50%) as white solid. LC-MS [M+H]312.4 (C₁₉H₂₁NO₃+H, requires 312.39).

Synthesis of(2S,3R)-3-hydroxy-2-[4-(4-morpholin-4-ylmethyl-benzyl)-benzoylamino]-butyricacid methyl ester (4)

A solution of compound (3) (31 mg, 0.09 mmol), HATU (34 g, 0.09 mmol)and DIEA (63 μl, 0.36 mmol) in DMF (800 μl) was maintained at ambienttemperature for 10 min followed by the addition of H-Thr-OMehydrochloride (18 mg, 0.11 mmol). Reaction mixture was stirred atambient temperature overnight followed by the dilution with EtOAc (100ml). Solution was extracted with water (20 ml×2) and brine (20 ml).Organic layer was dried over MgSO₄ and evaporated. Residue was dried invacuum to provide target compound (4) (31 mg, 82%) as yellow solid.LC-MS [M+H] 427.1 (C₂₄H₃₀N₂O₅+H, requires 427.53).

Synthesis ofN-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-benzyl)-benzamide(48-5)

A solution of hydroxylamine hydrochloride (15 mg, 0.22 mmol) in MeOH (2ml) was cooled to −5° C. followed by the addition of 25% NaOMe/MeOH (75μl, 0.33 mmol) under nitrogen. Reaction mixture was stirred at −5° C.for additional 5 min, cooled to −20° C., and solution of compound 4 (31mg, 0.07 mmol) in THF/MeOH (1:1, 2 ml) was added dropwise over theperiod of 5 min. Temperature of reaction mixture was raised to ambient.Completion of the reaction was monitored by LC-MS. Reaction mixture wasacidified with 1 N aq. HCl to pH˜7 and evaporated in vacuum. Residue wasdissolved in DMSO (500 μl) and subjected to HPLC purification.[Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flow rate=20 ml/min;mobile phase A: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1%TFA; gradient elution from 8% B to 28% B in 60 min., detection 254 nm].Fractions containing the desired product were combined and lyophylisedto provide trifluoroacetic salt of target product (48-5) (18.8 mg, 50%)as white solid. LC-MS [M+H] 428.1 (C₂₃H₂₉N₃O₅+H, requires 428.51).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (48-5) 0.07 18.8 50 99.3 428.1 2.73 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 mm, detection 254 nm]

Example 49

Examples 50A and 50B

Example 51

Example 52

Example 53

Example 54

Example 55

Example 56

Example 57

Example 58

Example 59(S)-3-amino-N-hydroxy-2-(7-((4-(morpholinomethyl)phenyl)ethynyl)-4-oxoquinazolin-3(4H)-yl)propanamide(59-8)

2-amino-4-iodobenzoic acid (2)

A solution of 2-amino-4-bromobenzoic acid (1) (2.15 g, 10 mmol) indegassed DMF (25 ml), CuI (3.80 g, 20 mmol) and KI (8.8 g, 50 mmol) washeated in a sealed tube at 130° C. for 18 h. The reaction mixture wascooled to room temperature and solid was filtered off. The filtrate wasconcentrated to dryness and the residue was stirred in ethyl acetate(400 ml) and water (100 ml) and the organic layer was separated andconcentrated. The crude product was then purified over a silica gelcolumn to give (2) (0.98 g).

(S)-methyl2-(2-amino-4-iodobenzamido)-3-(tert-butoxycarbonylamino)propanoate (4)

To a solution of 2-amino-4-iodobenzoic acid (2) (0.98 g, 3.37 mmol), DAPhydrochloride methyl ester (compound 3) (0.952 g, 3.74 mmol), DIEA (2mL, excess) in acetonitrile (26 mL) was added HATU (1.42 g, 3.74 mmol).The reaction mixture was stirred for 30 min and concentrated. The crudeproduct extracted with ethyl acetate (200 mL) and concentrated to give(4) and used as such in the next step.

(S)-methyl-3-(tert-butoxycarbonylamino)-2-(7-iodo-4-oxoquinazolin-3(4H)-yl)propanoate(5)

Compound (4) (1.42 g) was heated in neat trietyl orthoformate (18 mL) at120° C. for 24 h. Excess triethyl orthoformate was removed under reducedpressure and crude product was purified on silica gel column to givecompound (5) (0.48 g).

(S)-methyl-3-(tert-butoxycarbonylamino)-2-(7-((4-(morpholinomethyl)phenyl)ethynyl)-4-oxoquinazolin-3(4H)-yl)propanoate(7)

CuI (15 mg) and PdCl₂(PPh₃)₂ (21 mg) was added to a degassed THF (10 ml)solution of(S)-methyl-3-(tert-butoxycarbonylamino)-2-(7-iodo-4-oxoquinazolin-3(4H)-yl)propanoate(5) (0.48 g, 1.01 mmol), diisopropyl amine (2 mL) and4-(4-ethynylbenzyl)morpholine (6) (0.21 g, 1.05 mmol) and stirred for 1h. The solvent was removed and extracted with ethyl acetate (2×100 ml),dried over Na₂SO₄, and concentrated to give compound (7) (0.521 g).

(S)-3-amino-N-hydroxy-2-(7-((4-(morpholinomethyl)phenyl)ethynyl)-4-oxoquinazolin-3(4H)-yl)propanamide(59-8)

To a stirred solution of compound (7) (0.18 g, 0.338 mmol) andhydroxylamine hydrochloride (118 mg, 1.69 mmol) in methanol (10 ml), KOH(113 mg, 2 mmol) was added and stirred for 30 min. Excess solvent wasremoved and crude product was acidified with dilute AcOH (10%) andextracted with ethyl acetate (2×50 ml), dried and concentrated. Afterdeprotection of BOC group using HCl gas in ethyl acetate, crude productwas purified using HPLC to give compound (59-8). [M+H⁺]=448

Example 60N-Hydroxy-2-isopropyl-N′-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-phenyl]-malonamide

Synthesis of 2-(4-Iodo-phenylcarbamoyl)-3-methyl-butyric acid (1)

A solution of isopropylmalonic acid (613 mg, 4.2 mmol) and DIC (330 μl,2.1 mmol) in chloroform (2 ml) was maintained at ambient temperature for10 min followed by the addition of 4-iodoaniline (438 mg, 2.0 mmol).Reaction mixture was stirred at ambient temperature for additional 2 hand diluted with EtOAc (150 ml). Solution was extracted with water (30ml×2) and brine (30 ml), and dried over MgSO₄ (anh). Solvent wasevaporated in vacuum. Residue was dissolved in DCM (2 ml) and subjectedto flash chromatography on CombiFlash® Companion unit equipped withRediSep® flash column (normal phase, 35-60 micron average particle sizesilicagel, 80 g, Teledyne Isco); flow rate=60 ml/min; injection volume2.5 ml; mobile phase A: DCM; mobile phase B: MeOH; gradient 0-60% B in60 min. Fractions containing the desired product were combined andconcentrated in vacuum. Residue was dried in vacuum overnight to obtaintarget product (1) (89 mg, 13%) as white solid. LC-MS [M+H] 348.1(C₁₂H₁₄1 NO₃+H, requires 348.16).

Synthesis of 2-(4-iodo-phenylcarbamoyl)-3-methyl-butyric acid methylester (2)

A solution of compound (1) (89 g, 0.26 mmol) and HCl (conc., 500 μl) in2,2-dimethoxypropane (2.0 ml, 16 mmol) was maintained at ambienttemperature overnight. Reaction mixture was evaporated in vacuum.Residue was dissolved in i-PrOH (10 ml) and evaporated in vacuum. Theaforementioned procedure was repeated twice. Residue was triturated withether, filtrated, washed with ether and dried in vacuum overnight toobtained hydrochloric salt of target product (2) (84 mg, 99%) as whiteamorphous solid. LC-MS [M+H] 361.9 (C₁₃H₁₆1 NO₃+H, requires 362.19).

Synthesis of2-[4-(4-formyl-phenylethynyl)-phenylcarbamoyl]-3-methyl-butyric acidmethyl ester (3)

A solution of compound (2) (84 mg, 0.23 mmol), 4-ethynylbenzaldehyde (36mg, 0.28), Ph₃P (6 mg, 0.23 mmol) and di-isopropylamine (400 in THF(anh., 800 μl) was purged with dry nitrogen for 2 min followed by theaddition of mixture of PdCl₂(PPh₃)₂ (5 mg, 0.007 mmol) and CuI (3 mg,0.014 mmol). Reaction mixture was irradiated in microwave oven (max.power 250 W, 120° C.) for 10 min and cooled to ambient temperature.Solvents were evaporated in vacuum. Residue was dissolved in EtOAc (80ml) and extracted with 5% aq. NaHCO₃ (20 ml×2) and brine (20 ml). Theorganic layer was dried over anh. MgSO₄ and evaporated in vacuum.Residue was dried in vacuum overnight at ambient temperature to providetarget material (3) (82 mg, 99%) as brown solid. LC-MS [M+H] 364.2(C₂₂H₂₁NO₄+H, requires 364.43).

Synthesis of3-methyl-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-phenylcarbamoyl]-butyricacid methyl ester (4)

A solution of compound (3) (82 mg, 0.23 mmol), AcOH (52 μl, 0.92 mmol)and morpholine (30 μl, 0.35 mmol) in DCE (1 ml) was maintained atambient temperature for 10 min followed by the addition of sodiumtriacetoxyborohydride (98 mg, 0.46 mmol). Reaction mixture was stirredfor 2 h at ambient temperature. Reaction was quenched with 5% aq. NaHCO₃(20 ml) and extracted with EtOAc (50 ml×2). Organic layer was washedwith brine (20 ml), dried over anh. MgSO₄ and evaporated in vacuum.Residue was dried in vacuum overnight to provide target material (4) (98g, 98%) as yellowish solid. LC-MS [M+H] 435.0 (C₂₆H₃₀N₂O₄+H, requires435.55).

Synthesis ofN-hydroxy-2-isopropyl-N′-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-phenyl]-malonamide(60-5)

A solution of hydroxylamine hydrochloride (97 mg, 1.38 mmol) in MeOH(anh, 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (473 μl, 1.8 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (4) (98 mg, 0.23 mmol) in THF/MeOH (1:1, 2 ml) was addeddropwise over the period of 5 min. Temperature of reaction mixture wasraised to ambient. Completion of the reaction was monitored by LC-MS.After completion reaction mixture was acidified with 1 N aq. HCl to pH˜7and evaporated in vacuum. Residue was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophylised to provide trifluoroacetic salt of targetproduct (60-5) (12 mg, 9.4%) as white solid. LC-MS [M+H] 436.0(C₂₅H₂₉N₃O₄+H, requires 436.54).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (60-5) 0.23 12 9.4 99.8 436.0 3.31 *[Chromolith SpeedRodRP-18e C18 column (4.6 x 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm]

Example 61

Example 62 Synthesis of(R)—N-Hydroxy-2-[4-(4-morpholin-4-yl-methyl-phenylethynyl)-benzenesulfonylamino]-3-phenylpropionamide

Synthesis of 4-(4-bromo-benzyl)-morpholine (1)

To a solution of morpholine (2.61 ml, 30 mmol) in DMSO (25 ml) was added4-bromobenzylbromide (2.5 g, 10 mmol). Reaction mixture was stirred atambient temperature for 30 min. Solvents were evaporated in vacuum.Residue was dissolved in EtOAc (150 ml) and extracted with 5% aq. NaHCO₃(50 ml×2) and brine (50 ml). The organic layer was dried over anh. MgSO₄and evaporated in vacuum. Residue was dried in vacuum overnight atambient temperature to provide target material (1) (2.27 g, 87%) as awhite solid.

Synthesis of 4-(4-trimethylsilanylethynyl-benzyl)-morpholine (2)

To a solution of compound (1) (2.27 g, 8.9 mmol), ethynyltrimethylsilane(1.0 g, 10 mmol), PPh₃ (262 mg, 1 mmol) and dipropylamine (5 ml, 36.5mmol) in DMF (10 ml) was added a mixture of PdCl₂(PPh₃)₂ (130 mg, 0.18mmol) and CuI (100 mg, 0.5 mmol). Reaction mixture was irradiated inmicrowave oven (max. power 250 W, 120° C.) for 25 min and cooled toambient temperature. Reaction mixture was diluted with water (100 ml)and extracted with EtOAc (50 ml×2). Organic layer was washed with 5% aq.NaHCO₃ (50 ml) and brine (50 ml), and dried over MgSO₄ (anh). Solventwas evaporated in vacuum. Residue was dried in vacuum overnight toproduce target material (2) (2.36 g, 97%) as brown oil. LC-MS [M+H]274.1 (C₁₆H₂₃NOSi+H, requires 274.46).

4-(4-Ethynyl-benzyl)-morpholine (3)

To a solution of compound (2) (2.36 g, 8.6 mmol) in DCM (3 ml) was addedTFA (10 ml, 135 mmol). Reaction mixture was maintained at ambienttemperature for 1.5 h. Solvent was evaporated in vacuum. Residue wasdissolved in DMSO (15 ml) and subjected to HPLC purification.[Nanosyn-Pack Microsorb 100-10 C-18 column (50×300 mm); flow rate=50ml/min; injection volume 18 ml; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 5% B to 20% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and concentrated in vacuum. Residue was dissolved in ether(100 ml) and washed with 1 M aq. NaOH (50 ml), and brine (50 ml).Organic layer was dried over MgSO₄ (anh). Solvent was evaporated invacuum. Residue was dried in vacuum overnight to produce target material(3) (687 mg, 40%) as off-white solid. LC-MS [M+H] 202.4 (C₁₃H₁₅NO+H,requires 202.28).

Synthesis of(R)—N-Hydroxy-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzenesulfonylamino]-3-phenyl-propionamide (62-4)

2-Chlorotrityl-N-Fmoc-hydroxylamine resin (50 mg, 0.03 mmol) was placedin 3 cc plastic syringe equipped with plunger and plastic frit. Resinwas swelled in DCM (2 ml) at ambient temperature for 10 min. DCM wasdischarged followed by the treatment with 20% piperidine/DMF (2 ml×2×10min). Solvents were discharged and resin was washed with DCM (2 ml×6×5min) followed by the addition of mixture of Fmoc-D-Phe-OH (47 mg, 0.12mmol), HATU (46 mg, 0.12 mmol) and DIEA (42 μl, 0.24 mmol) in DMF (500μl). Syringe was agitating at 85 rpm on orbital shaker at ambienttemperature overnight. Solution was discharged and resin was washed withDMF (2 ml×6×5 min) followed by the treatment with 20% piperidine/DMF (2ml×2×10 min). Solvents were discharged and resin was washed with DCM (2ml×6×5 min) followed by the addition of mixture of compound (3) (48 mg,0.24 mmol), PdCl₂(PPh₃)₂ (20 mg, 0.03 mmol), CuI (10 mg, 0.05 mmol) anddi-propylamine (200 μl) in DMF (400 μl). Syringe was agitating at 85 rpmon orbital shaker at ambient temperature overnight. Solution wasdischarged and resin was washed with DMF (3 ml×4×5 min) and DCM (3ml×4×5 min) followed by the addition of 80% aq. TFA (2 ml). Syringe wasagitating at 85 rpm on orbital shaker at ambient temperature for 3 min.Solution was collected and evaporated in vacuum. Residue was dissolvedin DMSO (500 μl) and subjected to HPLC purification. [Phenomenex GeminiC-18 column, 110 Å (30×100 mm); flow rate=20 ml/min; mobile phase A:100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1% TFA; gradientelution from 8% B to 28% B in 60 min., detection 254 nm]. Fractionscontaining the desired product were combined and lyophylised to providetrifluoroacetic salt of target product (62-4) as white solid. LC-MS[M+H] 520.5 (C₂₈H₂₉N₃O₅S+H requires 520.63).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (62-4) 0.03 4.04 21 100 520.5 3.70 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.5 min, detection 254 nm]The following compounds were synthesized as described in this Example.

Compound # Structure [M + H] (62-5) 

550 (62-6) 

520 (62-7) 

474 (62-8) 

484 (62-9) 

490 (62-10)

470 (62-11)

456

Example 63(2S,3R)-3,N-Dihydroxy-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzenesulfinylamino]-butyramide

Synthesis of (2S,3R)-2-(4-bromo-benzenesulfinylamino)-3-hydroxy-butyricacid methyl ester (1)

A solution of 4-Bromobenzenesulfinic acid sodium salt dihydrate (250 mg,0.9 mmol) in SOCl₂ (3 ml, 41.1 mmol) was maintained at ambienttemperature for 3 h. Solvent was evaporated. Residue was dissolved inDCM (8 ml) and combined with H-Thr-OMe hydrochloride (253 mg, 1.5 mmol)followed by the dropwise addition of DIEA (500 μl, 3.0 mmol) over theperiod of 5 min. After stirring at ambient temperature for additionalhour reaction mixture was subjected to flash chromatography onCombiFlash® Companion unit equipped with RediSep® flash column (normalphase, 35-60 micron average particle size silicagel, 4 g, TeledyneIsco); flow rate=18 ml/min; injection volume 10 ml; mobile phase A:hexane; mobile phase B: EtOAc; gradient 0-100% B in 40 min. Fractionscontaining the desired product were combined and concentrated in vacuum.Residue was dried in vacuum overnight to provide target product (1) (100mg, 33%) as white solid. LC-MS [M+H] 337.8 (C₁₁H₁₄BrNO₄S+H, requires337.22).

Synthesis of(2S,3R)-2-[4-(4-Formyl-phenylethynyl)-benzenesulfinylamino]-3-hydroxy-butyricacid methyl ester (2)

A solution of compound (1) (100 mg, 0.30 mmol), 4-ethynylbenzaldehyde(43 mg, 0.32 mmol), Ph₃P (123 mg, 0.34) and di-propylamine (1 ml) in DMF(2 ml) was purged for 5 min with dry nitrogen. Catalysts PdCl₂(PPh₃)₂(30 mg, 0.043 mmol), and CuI (15 mg, 0.079 mmol) were added and reactionmixture was subjected to microwave irradiation (max. power 250 W, 120°C.) for 15 min. Reaction mixture was cooled to ambient temperature,diluted with EtOAc (100 ml) and extracted with water (30 ml), 2% aq.H₂SO₄ (30 ml), water (30 ml×2) and brine (30 ml). Organic layer wasdried over Na₂SO₄ and evaporated. Residue was dried in vacuum to providetarget compound (2) as brownish amorphous solid. LC-MS [M+H] 385.6(C₂₀H₁₉NO₅S+H, requires 386.45). Compound (2) was used as is for thenext transformation.

Synthesis of(2S,3R)-3-Hydroxy-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzenesulfinylamino]-butyricacid methyl ester (3)

A solution of compound (2) (0.30 mmol) and morpholine (52 μl, 0.6 mmol)in chloroform (10 ml) was treated with NaBH(OAc)₃ (100 mg, 0.47 mmol)for 5 h at ambient temperature under nitrogen. Reaction was quenchedwith 5% aq. NaHCO₃ (20 ml) and extracted with EtOAc (50 ml×2). Organiclayer was dried over anhydrous Na₂SO₄ and evaporated in vacuum. Residuewas dissolved in DCM (2 ml) and subjected to flash chromatography onCombiFlash® Companion unit equipped with RediSep® flash column (normalphase, 35-60 micron average particle size silicagel, 12 g, TeledyneIsco); flow rate=30 ml/min; injection volume 3 ml; mobile phase A: DCM;mobile phase B: MeOH; gradient 0-5% B in 15 min. Fractions containingthe desired product were combined and concentrated in vacuum. Residuewas dried in vacuum overnight to provide target product (3) (92 mg, 33%)as colorless oil. LC-MS [M+H] 457.3 (C₂₄H₂₈N₂O₅S+H, requires 457.58).

Synthesis of(2S,3R)-3,N-dihydroxy-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzenesulfinylamino]-butyramide(4)

A solution of hydroxylamine hydrochloride (84 mg, 1.2 mmol) in MeOH(anh., 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (411 μl, 3.15 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (3) (92 mg, 0.20 mmol) in THF/MeOH (1:1, 2 ml) was addeddropwise over the period of 5 min. Temperature of reaction mixture wasraised to ambient. Completion of the reaction was monitored by LC-MS.After completion reaction mixture was acidified with 1 N aq. HCl to pH˜7and evaporated in vacuum. Residue was dissolved in DMSO (800 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and neutralized with 1 N aq. NaOH. Organics wereevaporated in vacuum. Water layer was extracted with EtOAc (10 ml),dried over Na₂SO₄ and evaporated in vacuum. Residue was dried in vacuumovernight to provide free base of target product (4) (4 mg, 4.4%) aswhite solid. LC-MS [M+H] 458.1 (C₂₃H₂₇N₃O₅S+H, requires 458.56).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (4) 0.2 4 4.4 98.5 475.1 2.96 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.5 min, detection 254 nm

Example 64N-(Hydroxycarbamoyl-pyrrolidin-3-yl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide

Synthesis of 4-(4-formyl-phenylethynyl)-benzonitrile (1)

To a solution of 4-ethynylbenzonitrile (2.27 g, 17.9 mmol),4-iodobenzaldehyde (5.40 g, 23.3) and triethylamine (7.0 ml, 2.3 mmol)in THF (anh., 360 ml) was added mixture of PdCl₂(PPh₃)₂ (400 mg, 0.6mmol) and CuI (216 mg, 1.1 mmol). Reaction mixture was stirred atambient temperature overnight. Solvents were evaporated in vacuum.Residue was dissolved in EtOAc (150 ml) and extracted with 5% aq. NaHCO₃(50 ml×2) and brine (50 ml). The organic layer was dried over anh.Na₂SO₄ and evaporated in vacuum. Residue was dried in vacuum overnightat ambient temperature to provide target material (1) (3.75 g, 90%) asyellow solid.

Synthesis of 4-(4-morpholin-4-ylmethylphenylethynyl)-benzonitrile (2)

To a solution of compound (1) (2.0 g, 8.6 mmol) and morpholine (1.08 ml,12.4 mmol) in chloroform (30 ml) was added sodium triacetoxyborohydride(2.80 g, 13.2 mmol). Reaction mixture was stirred for 4 h at ambienttemperature. Reaction was quenched with 5% aq. NaHCO₃ (50 ml) andextracted with EtOAc (150 ml×2). Organic layer was washed with brine (50ml), dried over anh. Na₂SO₄ and evaporated in vacuum. Residue was driedin vacuum overnight to provide target material (2)(2.65 g, 97%) asoff-white solid. LC-MS [M+H] 303.2 (C₂₀H₁₈N₂O+H, requires 303.39).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoic acid (3)

A mixture of compound (2) (2.65 g, 8.8 mmol) and 2M aq. LiOH (22 ml, 44mmol) in dioxane (20 ml) was irradiated in microwave oven (max. power250 W, 130° C.) for 45 min and cooled to ambient temperature. Reactionmixture was diluted with water (300 ml) and extracted with EtOAc (100ml×2). Water layer was acidified with 1 M HCl to pH˜3 and extracted withEtOAc (300 ml×3). Organic layer was washed with water (50 ml×2) andbrine (50 ml), and dried over MgSO₄ (anh). Solvent was evaporated invacuum. Residue was dried in vacuum overnight to produce hydrochloricsalt of target material (3) (2.23 g, 71%) as yellowish solid. LC-MS[M+H] 322.1 (C₂₀H₁₉NO₃+H, requires 322.39).

Synthesis of3-(tert-butoxycarbonylamino-carboxy-methyl)-pyrrolidine-1-carboxylicacid 9H-fluoren-9-ylmethyl ester (4)

To a solution of Boc-DL-pyrrolidin-3-yl-acetic acid (2.0 g, 8.6 mmol) inwater (2 ml) was added solution of Fmoc-OSu (621 mg, 1.84 mmol) in ACN(5 ml). Reaction mixture was stirred for 2 h at ambient temperature.Reaction was diluted with EtOAc (50 ml). Organic layer was washed withwater (30 ml×2) and brine (30 ml), dried over anh. Na₂SO₄ and evaporatedin vacuum. Residue was dried in vacuum overnight to provide targetmaterial (4) (500 mg, 58%) as white solid. LC-MS [M+H] 467.2(C₂₆H₃₀H₂O₆+H, requires 467.55).

Synthesis of 3-(amino-methoxycarbonyl-methyl)-pyrrolidine-1-carboxylicacid 9H-fluoren-9-ylmethyl ester (5)

A solution of compound (4) (500 mg, 1.07 mmol) and HCl (conc., 100 μl)in 2,2-dimethoxypropane (3 ml, 24 mmol) was maintained at ambienttemperature overnight. Reaction mixture was evaporated in vacuum.Residue was dissolved in DCM (1 ml) and subjected to flashchromatography on CombiFlash® Companion unit equipped with RediSep®flash column (normal phase, 35-60 micron average particle sizesilicagel, 12 g, Teledyne Isco); flow rate=30 ml/min; injection volume1.5 ml; mobile phase A: DCM; mobile phase B: MeOH; gradient 0-30% B in43 min. Fractions containing the desired product were combined andconcentrated in vacuum. Residue was triturated with ether, filtrated,washed with ether and dried in vacuum overnight to obtained hydrochloricsalt of target product (5) (300 mg, 67%) as off-white powder. LC-MS[M+H] 381.1 (C₂₂H₂₄N₂O₄+H, requires 381.46).

Synthesis of3-{methoxycarbonyl-[4-(4-morpholin-4-ylmethyl-phenylethynyl)benzoylamino]-methyl}-pyrrolidine-1-carboxylicacid 9H-fluoren-9-ylmethyl ester (6)

A solution of compound (3) hydrochloride (110 mg, 0.34 mmol), HATU (136mg, 0.36 mmol) and DIEA (330 μl, 1.9 mmol) in DMF (1.5 ml) wasmaintained at ambient temperature for 10 min followed by the addition ofcompound 5 hydrochloride (150 mg, 0.36 mmol). Reaction mixture wasstirred at ambient temperature overnight, diluted with EtOAc (80 ml) andextracted with water (20 ml×2) and brine (20 ml). Organic layer wasdried over anh. MgSO₄, evaporated in vacuum and dried in vacuumovernight to provide target product (6) (228 mg, 99%) as brown solid.LC-MS [M+H] 684.6 (C₄₂H₄₁N₃O₆+H, requires 684.82).

Synthesis ofN-(hydroxycarbamoyl-pyrrolidin-3-yl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(64-7)

A solution of hydroxylamine hydrochloride (140 mg, 0.33 mmol) in MeOH(anh, 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (678 μl, 2.0 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (6) (228 mg, 0.33 mmol) in THF/MeOH (1:1, 2 ml) was addeddropwise over the period of 5 min. Temperature of reaction mixture wasraised to ambient. Completion of the reaction was monitored by LC-MS.After completion reaction mixture was acidified with 1 N aq. HCl to pH˜7and evaporated in vacuum. Residue was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophylised to provide di-trifluoroacetic salt oftarget product (64-7) (4.8 mg, 2.1%) as white solid. LC-MS [M+H] 463.2(C₂₆H₃₀N₄O₄+H, requires 463.56).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (64-7) 0.33 4.8 2.1 97.4 463.2 2.31 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm]

Example 65N—((S)-Hydroxycarbamoyl-piperidin-4-yl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide

Synthesis of 4-(4-formyl-phenylethynyl)-benzonitrile (1)

To a solution of 4-ethynylbenzonitrile (2.27 g, 17.9 mmol),4-iodobenzaldehyde (5.40 g, 23.3) and triethylamine (7.0 ml, 2.3 mmol)in THF (anh., 360 ml) was added mixture of PdCl₂(PPh₃)₂ (400 mg, 0.6mmol) and CuI (216 mg, 1.1 mmol). Reaction mixture was stirred atambient temperature overnight. Solvents were evaporated in vacuum.Residue was dissolved in EtOAc (150 ml) and extracted with 5% aq. NaHCO₃(50 ml×2) and brine (50 ml). The organic layer was dried over anh.Na₂SO₄ and evaporated in vacuum. Residue was dried in vacuum overnightat ambient temperature to provide target material (1) (3.75 g, 90%) asyellow solid.

Synthesis of 4-(4-morpholin-4-ylmethylphenylethynyl)-benzonitrile (2)

To a solution of compound (1) (2.0 g, 8.6 mmol) and morpholine (1.08 ml,12.4 mmol) in chloroform (30 ml) was added sodium triacetoxyborohydride(2.80 g, 13.2 mmol). Reaction mixture was stirred for 4 h at ambienttemperature. Reaction was quenched with 5% aq. NaHCO₃ (50 ml) andextracted with EtOAc (150 ml×2). Organic layer was washed with brine (50ml), dried over anh. Na₂SO₄ and evaporated in vacuum. Residue was driedin vacuum overnight to provide target material (2) (2.65 g, 97%) asoff-white solid. LC-MS [M+H] 303.2 (C₂₀H₁₈N₂O+H, requires 303.39).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoic acid (3)

A mixture of compound (2) (2.65 g, 8.8 mmol) and 2M aq. LiOH (22 ml, 44mmol) in dioxane (20 ml) was irradiated in microwave oven (max. power250 W, 130° C.) for 45 min and cooled to ambient temperature. Reactionmixture was diluted with water (300 ml) and extracted with EtOAc (100ml×2). Water layer was acidified with 1 M HCl to pH˜3 and extracted withEtOAc (300 ml×3). Organic layer was washed with water (50 ml×2) andbrine (50 ml), and dried over MgSO₄ (anh). Solvent was evaporated invacuum. Residue was dried in vacuum overnight to produce hydrochloricsalt of target material (3) (2.23 g, 71%) as yellowish solid. LC-MS[M+H] 322.1 (C₂₀H₁₉NO₃+H, requires 322.39).

Synthesis of4-{(S)-carboxy-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-methyl}-piperidine-1-carboxylicacid tert-butyl ester (4)

A solution of compound (3) (110 mg, 0.31 mmol), HATU (136 mg, 0.36 mmol)and DIEA (330 μl, 1.9 mmol) in DMF (1.5 ml) was maintained at ambienttemperature for 30 min. A mixture of H-Gly[4-Pip(Boc)]-OH (150 mg, 0.36mmol) and BSA (170 μl, 0.7 mmol) in dioxane (1.5 ml) was stirred at 100°C. for 10 min and cooled to the ambient temperature. Reaction mixtureswere combined and stirred at ambient temperature overnight followed bythe dilution with EtOAc (100 ml). Solution was extracted with water (20ml), 2% aq. H₂SO₄ (20 ml), water (20 ml×2) and brine (20 ml). Organiclayer was dried over MgSO₄ and evaporated. Residue was dried in vacuumto provide target compound (4) (172 mg, 99%) as yellow solid. LC-MS[M+H] 562.2 (C₃₂H₃₉N₃O₆+H, requires 562.69).

Synthesis of(S)-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-piperidin-4-yl-aceticacid methyl ester (5)

A solution of compound (4) (172 mg, 0.31 mmol) and HCl (conc., 1 ml) in2,2-dimethoxypropane (5 ml, 40 mmol) was maintained at 60° C. for 3 h.Reaction mixture was cooled to ambient temperature and evaporated invacuum. Residue was dissolved in DCM (1 ml) and subjected to flashchromatography on CombiFlash® Companion unit equipped with RediSep flashcolumn (normal phase, 35-60 micron average particle size silicagel, 4 g,Teledyne Isco); flow rate=18 ml/min; injection volume 1.5 ml; mobilephase A: hexane; mobile phase B: EtOAc; gradient 0-20% B in 30 min.Fractions containing the desired product were combined and concentratedin vacuum. Residue was dried in vacuum overnight to obtainedhydrochloric salt of target product (5) (44 mg, 91%) as off-whitepowder. LC-MS [M+H] 476.4 (C₂₈H₃₃N₃O₄+H, requires 476.60).

Synthesis ofN—((S)-hydroxycarbamoyl-piperidin-4-yl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(65-6)

A solution of hydroxylamine hydrochloride (118 mg, 0.28 mmol) in MeOH(anh, 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (575 μl, 2.52 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (5) (144 mg, 0.28 mmol) in THF/MeOH (1:1, 2 ml) was addeddropwise over the period of 5 min. Temperature of reaction mixture wasraised to ambient. Completion of the reaction was monitored by LC-MS.After completion reaction mixture was acidified with 1 N aq. HCl to pH˜7and evaporated in vacuum. Residue was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophylised to provide di-trifluoroacetic salt oftarget product (65-6) (11.1 mg, 5.6%) as white solid. LC-MS [M+H] 477.5(C₂₆H₃₀N₄O₄+H, requires 463.56).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (65-6) 0.28 11.1 5.6 99.5 477.5 2.36 *[ChromolithSpeedRod RP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobilephase A: 0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elutionfrom 5% B to 100% B over 9.6 min, detection 254 nm]

Example 66N—((S)-Hydroxycarbamoyl-phenyl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide

Synthesis of 4-(4-formyl-phenylethynyl)-benzonitrile (1)

To a solution of 4-ethynylbenzonitrile (2.27 g, 17.9 mmol),4-iodobenzaldehyde (5.40 g, 23.3) and triethylamine (7.0 ml, 2.3 mmol)in THF (anh., 360 ml) was added mixture of PdCl₂(PPh₃)₂ (400 mg, 0.6mmol) and CuI (216 mg, 1.1 mmol). Reaction mixture was stirred atambient temperature overnight. Solvents were evaporated in vacuum.Residue was dissolved in EtOAc (150 ml) and extracted with 5% aq. NaHCO₃(50 ml×2) and brine (50 ml). The organic layer was dried over anh.Na₂SO₄ and evaporated in vacuum. Residue was dried in vacuum overnightat ambient temperature to provide target material (1) (3.75 g, 90%) asyellow solid.

Synthesis of 4-(4-morpholin-4-ylmethylphenylethynyl)-benzonitrile (2)

To a solution of compound (1) (2.0 g, 8.6 mmol) and morpholine (1.08 ml,12.4 mmol) in chloroform (30 ml) was added sodium triacetoxyborohydride(2.80 g, 13.2 mmol). Reaction mixture was stirred for 4 h at ambienttemperature. Reaction was quenched with 5% aq. NaHCO₃ (50 ml) andextracted with EtOAc (150 ml×2). Organic layer was washed with brine (50ml), dried over anh. Na₂SO₄ and evaporated in vacuum. Residue was driedin vacuum overnight to provide target material (2) (2.65 g, 97%) asoff-white solid. LC-MS [M+H] 303.2 (C₂₀H₁₈N₂O+H, requires 303.39).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoic acid

A mixture of compound (2) (2.65 g, 8.8 mmol) and 2M aq. LiOH (22 ml, 44mmol) in dioxane (20 ml) was irradiated in microwave oven (max. power250 W, 130° C.) for 45 min and cooled to ambient temperature. Reactionmixture was diluted with water (300 ml) and extracted with EtOAc (100ml×2). Water layer was acidified with 1 M HCl to pH˜3 and extracted withEtOAc (300 ml×3). Organic layer was washed with water (50 ml×2) andbrine (50 ml), and dried over MgSO₄ (anh). Solvent was evaporated invacuum. Residue was dried in vacuum overnight to produce hydrochloricsalt of target material (3) (2.23 g, 71%) as yellowish solid. LC-MS[M+H] 322.1 (C₂₀H₁₉NO₃+H, requires 322.39).

Synthesis ofS)-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-phenyl-aceticacid methyl ester (4)

A solution of compound (3) hydrochloride (143 mg, 0.4 mmol), HATU (152mg, 0.4 mmol) and DIEA (278 μl, 1.6 mmol) in DMF (1.5 ml) was maintainedat ambient temperature for 10 min followed by the addition of H-Phg-OMehydrochloride (89 mg, 0.44 mmol). Reaction mixture was stirred atambient temperature overnight, diluted with EtOAc (80 ml) and extractedwith water (20 ml×2) and brine (20 ml). Organic layer was dried overanh. MgSO₄, evaporated in vacuum and dried in vacuum overnight toprovide target product (4) (181 mg, 97%) as brown amorphous solid. LC-MS[M+H] 469.2 (C₂₉H₂₈N₂O₄+H, requires 469.57).

Synthesis ofN—((S)-hydroxycarbamoyl-phenyl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(66-5)

A solution of hydroxylamine hydrochloride (168 mg, 2.4 mmol) in MeOH(anh, 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (832 μl, 3.78 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (4) (181 mg, 0.4 mmol) in THF/MeOH (1:1, 2 ml) was addeddropwise over the period of 5 min. Temperature of reaction mixture wasraised to ambient. Completion of the reaction was monitored by LC-MS.After completion reaction mixture was acidified with 1 N aq. HCl to pH˜7and evaporated in vacuum. Residue was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophylised to provide trifluoroacetic salt of targetproduct (66-5) (143 mg, 8.3%) as white solid. LC-MS [M+H] 470.2(C₂₈H₂₇N₃O₄+H, requires 470.56).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (66-5) 0.4 143 61 99.4 470.2 3.46 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1 %TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm]The following compounds were synthesized as described in this Example.

Compound # Structure [M + H] (66-6)

Example 67N-[Hydroxycarbamoyl-(tetrahydro-pyran-4-yl)-methyl]-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide

Synthesis of 4-(4-formyl-phenylethynyl)-benzonitrile (1)

To a solution of 4-ethynylbenzonitrile (2.27 g, 17.9 mmol),4-iodobenzaldehyde (5.40 g, 23.3) and triethylamine (7.0 ml, 2.3 mmol)in THF (anh., 360 ml) was added mixture of PdCl₂(PPh₃)₂ (400 mg, 0.6mmol) and CuI (216 mg, 1.1 mmol). Reaction mixture was stirred atambient temperature overnight. Solvents were evaporated in vacuum.Residue was dissolved in EtOAc (150 ml) and extracted with 5% aq. NaHCO₃(50 ml×2) and brine (50 ml). The organic layer was dried over anh.Na₂SO₄ and evaporated in vacuum. Residue was dried in vacuum overnightat ambient temperature to provide target material (1) (3.75 g, 90%) asyellow solid.

Synthesis of 4-(4-morpholin-4-ylmethylphenylethynyl)-benzonitrile (2)

To a solution of compound (1) (2.0 g, 8.6 mmol) and morpholine (1.08 ml,12.4 mmol) in chloroform (30 ml) was added sodium triacetoxyborohydride(2.80 g, 13.2 mmol). Reaction mixture was stirred for 4 h at ambienttemperature. Reaction was quenched with 5% aq. NaHCO₃ (50 ml) andextracted with EtOAc (150 ml×2). Organic layer was washed with brine (50ml), dried over anh. Na₂SO₄ and evaporated in vacuum. Residue was driedin vacuum overnight to provide target material (2) (2.65 g, 97%) asoff-white solid. LC-MS [M+H] 303.2 (C₂₀H₁₈N₂O+H, requires 303.39).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoic acid (3)

A mixture of compound (2) (2.65 g, 8.8 mmol) and 2M aq. LiOH (22 ml, 44mmol) in dioxane (20 ml) was irradiated in microwave oven (max. power250 W, 130° C.) for 45 min and cooled to ambient temperature. Reactionmixture was diluted with water (300 ml) and extracted with EtOAc (100ml×2). Water layer was acidified with 1 M HCl to pH˜3 and extracted withEtOAc (300 ml×3). Organic layer was washed with water (50 ml×2) andbrine (50 ml), and dried over MgSO₄ (anh). Solvent was evaporated invacuum. Residue was dried in vacuum overnight to produce hydrochloricsalt of target material (3) (2.23 g, 71%) as yellowish solid. LC-MS[M+H] 322.1 (C₂₀H₁₉NO₃+H, requires 322.39).

Synthesis of[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-(tetrahydro-pyran-4-yl)-aceticacid methyl ester (4)

A solution of compound (3) hydrochloride (110 mg, 0.31 mmol), HATU (167mg, 0.44 mmol) and DIEA (330 μl, 2.0 mmol) in DMF (3 ml) was maintainedat ambient temperature for 10 min followed by the addition ofDL-(4-tetrahydropyranyl)Ala-OMe hydrochloride (84 mg, 0.4 mmol).Reaction mixture was stirred at ambient temperature overnight, dilutedwith EtOAc (100 ml) and extracted with water (30 ml×2) and brine (30ml). Organic layer was dried over anh. MgSO₄, evaporated in vacuum anddried in vacuum overnight to provide target product (4) (144 mg, 98%) asbrown solid. LC-MS [M+H] 477.2 (C₂₈H₃₂N₂O₅+H, requires 477.59).

Synthesis ofN-[hydroxycarbamoyl-(tetrahydro-pyran-4-yl)-methyl]-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(67-5)

A solution of hydroxylamine hydrochloride (126 mg, 1.8 mmol) in MeOH(anh, 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (616 μl, 2.7 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (4) (144 mg, 0.30 mmol) in THF/MeOH (1:1, 2 ml) was addeddropwise over the period of 5 min. Temperature of reaction mixture wasraised to ambient. Completion of the reaction was monitored by LC-MS.After completion reaction mixture was acidified with 1 N aq. HCl to pH˜7and evaporated in vacuum. Residue was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophylised to provide trifluoroacetic salt of targetproduct (67-5) (65.7 mg, 37%) as white solid. LC-MS [M+H] 408.2(C₂₇H₃₁H₃O₅+H, requires 478.58).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (67-5) 0.30 65.7 37 99.9 478.3 2.85 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 mm, detection 254 nm]The following compounds were synthesized as described in this Example.

Compound # Structure [M + H] (67-7)

476.3 (67-8)

476.2 (67-9)

526.2

Example 68N—((S)-Cyclopropyl-hydroxycarbamoyl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide

Synthesis of 4-(4-formyl-phenylethynyl)-benzonitrile (1)

To a solution of 4-ethynylbenzonitrile (2.27 g, 17.9 mmol),4-iodobenzaldehyde (5.40 g, 23.3) and triethylamine (7.0 ml, 2.3 mmol)in THF (anh., 360 ml) was added mixture of PdCl₂(PPh₃)₂ (400 mg, 0.6mmol) and CuI (216 mg, 1.1 mmol). Reaction mixture was stirred atambient temperature overnight. Solvents were evaporated in vacuum.Residue was dissolved in EtOAc (150 ml) and extracted with 5% aq. NaHCO₃(50 ml×2) and brine (50 ml). The organic layer was dried over anh.Na₂SO₄ and evaporated in vacuum. Residue was dried in vacuum overnightat ambient temperature to provide target material (1) (3.75 g, 90%) asyellow solid.

Synthesis of 4-(4-morpholin-4-ylmethylphenylethynyl)-benzonitrile (2)

To a solution of compound (1) (2.0 g, 8.6 mmol) and morpholine (1.08 ml,12.4 mmol) in chloroform (30 ml) was added sodium triacetoxyborohydride(2.80 g, 13.2 mmol). Reaction mixture was stirred for 4 h at ambienttemperature. Reaction was quenched with 5% aq. NaHCO₃ (50 ml) andextracted with EtOAc (150 ml×2). Organic layer was washed with brine (50ml), dried over anh. Na₂SO₄ and evaporated in vacuum. Residue was driedin vacuum overnight to provide target material (2) (2.65 g, 97%) asoff-white solid. LC-MS [M+H] 303.2 (C₂₀H₁₈N₂O+H, requires 303.39).

Synthesis of 4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoic acid (3)

A mixture of compound (2) (2.65 g, 8.8 mmol) and 2M aq. LiOH (22 ml, 44mmol) in dioxane (20 ml) was irradiated in microwave oven (max. power250 W, 130° C.) for 45 min and cooled to ambient temperature. Reactionmixture was diluted with water (300 ml) and extracted with EtOAc (100ml×2). Water layer was acidified with 1 M HCl to pH˜3 and extracted withEtOAc (300 ml×3). Organic layer was washed with water (50 ml×2) andbrine (50 ml), and dried over MgSO₄ (anh). Solvent was evaporated invacuum. Residue was dried in vacuum overnight to produce hydrochloricsalt of target material (3) (2.23 g, 71%) as yellowish solid. LC-MS[M+H] 322.1 (C₂₀H₁₉NO₃+H, requires 322.39).

Synthesis of (S)-amino-cyclopropyl-acetic acid methyl ester (4)

A solution of Boc-L-cyclopropylglycine (97 mg, 0.45 mmol) and HCl(conc., 500 μl) in 2,2-dimethoxypropane (5 ml, 40 mmol) was maintainedat 40° C. overnight. Reaction mixture was evaporated in vacuum. Residuewas dissolved in i-PrOH (10 ml) and evaporated in vacuum. Theaforementioned procedure was repeated twice. Residue was triturated withether, filtrated, washed with ether and dried in vacuum overnight toobtained hydrochloric salt of target product (4) (73 mg, 98%) asoff-white powder. LC-MS [M+H] 130.0 (C₆H₁₁NO₂+H, requires 130.17).

Synthesis of(S)-cyclopropyl-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-aceticacid methyl ester (5)

A solution of compound (3) hydrochloride (143 mg, 0.4 mmol), HATU (176mg, 0.44 mmol) and DIEA (313 μl, 1.8 mmol) in DMF (1 ml) was maintainedat ambient temperature for 10 min followed by the addition of compound(4) hydrochloride (73 mg, 0.44 mmol). Reaction mixture was stirred atambient temperature overnight, diluted with EtOAc (80 ml) and extractedwith water (20 ml×2) and brine (20 ml). Organic layer was dried overanh. MgSO₄ and evaporated in vacuum. Residue was dissolved in DCM (1 ml)and subjected to flash chromatography on CombiFlash® Companion unitequipped with RediSep® flash column (normal phase, 35-60 micron averageparticle size silicagel, 4 g, Teledyne Isco); flow rate=18 ml/min;injection volume 1.5 ml; mobile phase A: DCM; mobile phase B: MeOH;gradient 0-30% B in 38 min. Fractions containing the desired productwere combined and concentrated in vacuum. Residue was dried in vacuumovernight to obtain target product 5 (112 mg, 65%) as off-white powder.LC-MS [M+H] 433.2 (C₂₆H₂₈N₂O₄+H, requires 433.53).

Synthesis ofN—((S)-cyclopropyl-hydroxycarbamoyl-methyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(68-6)

A solution of hydroxylamine hydrochloride (107 mg, 1.6 mmol) in MeOH(anh, 2 ml) was cooled to −5° C. followed by the addition of 25%NaOMe/MeOH (534 μl, 2.34 mmol) under nitrogen. Reaction mixture wasstirred at −5° C. for additional 5 min, cooled to −20° C., and solutionof compound (5) (122 mg, 0.26 mmol) in THF/MeOH (1:1, 2 ml) was addeddropwise over the period of 5 min. Temperature of reaction mixture wasraised to ambient. Completion of the reaction was monitored by LC-MS.After completion reaction mixture was acidified with 1 N aq. HCl to pH˜7and evaporated in vacuum. Residue was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 ml/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophylised to provide trifluoroacetic salt of targetproduct (68-6) (39.5 mg, 28%) as white solid. LC-MS [M+H] 434.3(C₂₅H₂₇N₃O₄+H, requires 434.52).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* (68-6) 0.26 39.5 28 100 434.3 2.98 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm]

Example 69

Example 70

Example 71

Example 72

Example 73

Example 74

Example 75

Example 76

Example 77

Example 78

Example 79

Example 80

Example 81

General Procedures for Following Examples

Method 1-A (1-Bromo-1-alkyne/Alkyne Coupling)

A solution of the 1-bromo-1-alkyne (0.35 mmol, 1.0 equiv), alkyne (0.35mmol, 1.0 equiv), PdCl₂(PPh₃)₂ (0.0035 mmol, 0.01 equiv), P(tBu)₃ HBF₄(0.007 mmol, 0.02 equiv), Cu(I)I (0.0018 mmol, 0.005 equiv) and Et₃N(1.05 mmol, 3.0 equiv) in deoxygenated THF (5 mL) was allowed to stir atambient temperature under N₂ until the starting material was consumed asdetermined by TLC or LC-MS analysis. The solution was concentrated invacuo and purified by normal phase flash chromatography to provide thedesired compound.

Method 1-B (Vinyldibromide/Alkyne Coupling)

A solution of vinyldibromide (2.0 mmol, 1.0 equiv), alkyne* (2.8 mmol,1.4 equiv), Pd₂(dba)₃ (0.02 mmol, 0.01 equiv),tri(4-methoxyphenyl)phosphine (0.08 mmol, 0.04 equiv) and Et₃N (6.0mmol, 3.0 equiv) in deoxygenated DMF (5 mL) was heated at 80° C. for 2-6h under nitrogen. The reaction was monitored by LC-MS untildisappearance of the vinyldibromide. The reaction mixture was dilutedwith EtOAc (120-150 mL), washed with water (2×50 mL), brine (1×50 mL)dried (Na₂SO₄) and concentrated in vacuo. The crude material waspurified by normal phase flash chromatography to provide the desiredcompound. * In the case of a volatile alkyl- or cycloalkyl acetylene,amount of the acetylene was increased to 2-3 equivalents, and thereaction vessel was sealed before heating.

Method 1-C (Cadiot-Chodkiewicz Coupling)

To a solution of the alkyne (8.37 mmol, 1.0 equiv) in MeOH (25 mL) at 0°C. was added aqueous EtNH₂ (70%, 35 mL) and Cu(I)Cl (0.419 mmol, 0.05equiv). The this solution at 0° C. was then added a solution of the1-bromo- 1-alkyne (8.37 mmol, 1.0 equiv) in THF (25 mL). Solid NH₂OH HCl(8.37 mmol, 1.0 equiv) was then added in one portion and the solutionwas allowed to stir at 0° C. until starting material was consumed asdetermined by TLC or LC-MS analysis. The solution was then diluted withEtOAc (50 mL) and deionized water (50 mL). The layers were separated andthe organic layer was washed with aqueous HCl (0.1 N, 2×25 mL) and brine(1×25 mL). The organics were then dried (Na₂SO₄), filtered andconcentrated in vacuo. Crude material was then purified by flashchromatography to provide the desired compound.

Method 2-A (Basic Hydrolysis)

To a solution of the methyl ester (1.0-1.5 mmol, 1.0 equiv) in1,4-dioxane* (2-4 mL) was added 1.5 N NaOH (aq) (4-6 mL, 6.0 equiv) andthe mixture was heated at 70-90° C. for 1-4 h, monitored by LC-MS untilstarting material was consumed. After cooling to ambient temperature,the reaction mixture was acidified with 2 N HCl (aq)**, and the solidwas collected by filtration, washed with water (2×) and dried under highvacuum to give the desired compound. The product was used withoutfurther purification in the next synthetic step. * Alternatively, THF orEtOH can be used.** Alternatively, 10% H₃PO₄ (aq) or AcOH can be used.

Method 2-B (Acidic Hydrolysis)

A solution of the methyl ester (1-2 mmol, 1.0 equiv) in 2 N HCl (aq) (8mL, 8-16 equiv) was heated at 80° C. for 6-10 min, monitored by LC-MSuntil starting material was consumed. The reaction mixture was cooled to5° C. and the solid was collected by filtration, washed with water anddried under high vacuum to give the desired compound. The product wasused without further purification in the next synthetic step.

Method 3-A (HATU Coupling)

To a solution of the acid (0.50 mmol, 1.0 equiv) and DIEA (1.5 mmol, 3.0equiv) in DMF (0.1-0.5 M) was added HATU (0.60 mmol, 1.2 equiv), and themixture was stirred at ambient temperature* for 5 min. The amine (0.55mmol, 1.1 eq) was then added and the mixture was stirred at ambienttemperature for 18 h or until the starting material was consumed asdetermined by TLC or LC-MS analysis. The mixture was then added toaqueous HCl (0.1 N, 50 mL) and extracted with EtOAc (3×25 mL). Combinedorganic layers were washed with water (1×25 mL) and brine (1×25 mL),dried (Na₂SO₄), filtered and concentrated in vacuo to give the desiredcompound. Crude product may be used in next synthetic step withoutadditional purification or purified by normal phase flashchromatography. * An ice/water bath should be used if scale more 2 mmol.

Method 3-B (EDC/HOBT Coupling)

The carboxylic acid (1.15 mmol, 1.0 equiv), amine (1.27 mmol, 1.1equiv), EDC (2.30 mmol, 2.0 euiv) and HOBT (2.30 mmol, 2.0 equiv) wereslurried in anhydrous CH₂Cl₂ (12 mL). To this slurry was addedN,N-diisopropylethylamine (5.40 mmol, 4.0 equiv) in one portion. Thesolution was then allowed to stir at ambient temperature until completeconversion is observed by LCMS analysis. The solution was thenconcentrated in vacuo and the crude solid was purified by normal phaseflash chromatography to provide the desired compound.

Method 4-A (Boc Deprotection)

To the Boc protected amine (0.5 mmol, 1.0 equiv) was added 4 MHCl/dioxane (3-6 mL), and the mixture was stirred at ambient temperaturefor 0.5-2 h, or until complete as determined by TLC or LC-MS analysis.Volatiles were removed in vacuo or under a stream of nitrogen to givethe desired compound as the hydrochloride salt.

Method 4-B (Boc Deprotection)

To Boc protected amine (0.50 mmol, 1.0 equiv) in CH₂Cl₂ (10 mL) wasadded TFA (2 mL) and the mixture was stirred at ambient temperatureuntil complete as determined by TLC or LC-MS analysis. Volatiles wereremoved in vacuo or under a stream of nitrogen to give the desiredcompound as the TFA salt.

Method 4-C(Fmoc Deprotection)

To Fmoc protected amine (13.5 mmol, 1.0 equiv) in DMF (135 mL) at 0° C.was added morpholine (25 mL). The solution was allowed to stir at 0° C.until complete as determined by TLC or LC-MS analysis. The solution wasthen vacuum filtered through a pad of Celite and diluted with EtOAc (400mL). The organics were then washed with deionized water (3×100 mL),dried (MgSO₄), filtered and concentrated in vacuo. The crude materialwas then purified by normal phase flash chromatography.

Method 5-A (Hydroxamate Formation, Aqueous)

Isopropyl alcohol (1-4 mL) was added to a corresponding methyl ester(˜0.5 mmol, 1.0 eq) and the mixture was optionally cooled in anice/water bath for 5 min. NH₂OH (50% aq) (1-4 mL) was added to themixture, dropwise for the first ½ vol. After 5 min, the ice bath wasremoved and the reaction mixture was stirred for 6-24 h, or untilcomplete as determined by LC-MS analysis of the reaction mixture.Solvent volume was reduced by half using a nitrogen stream and water(10-15 mL) was added. The suspension was thoroughly agitated (vibromixer and sonication), centrifuged and the supernatant was discarded.Water (10-15 mL) was added to the solid and the suspension wasthoroughly agitated, centrifuged and the supernatant was discarded. Wetsolid was optionally dried by lyophilization to give the crudecorresponding hydroxamate.

Method 5-B (Hydroxamate Formation and Fmoc Deprotection, Anhydrous)

To a stirred suspension of a corresponding methyl ester (˜0.5 mmol, 1.0eq) and hydroxylamine hydrochloride (5 mmol, 10 eq) in anhydrous MeOH (2mL) and optionally anhyd THF (2 mL) cooled at −5° C. in a ice/salt/waterbath, was added dropwise 25% NaOMe/MeOH (1.2 mL, 10 mmol) undernitrogen. The reaction mixture was stirred at −5° C. for an additional 5min then stirred for 0.5-2 h or until complete as determined by LC-MSanalysis² of the reaction mixture. Reaction mixture was cooled,acidified to pH˜6 with 1 M HCl, and concentrated in vacuo or under astream of nitrogen to give the crude corresponding hydroxamate.

Method 6 (DAST Fluorination)

To a stirred solution of the corresponding alcohol (2.8 mmol, 1.0 equiv)in anhydrous CH₂Cl₂ (50 mL) was slowly added diethylamino sulfurtrifluoride (2.8 mmol, 1.0 equiv) at 0° C. The solution was allowed tostir at 0° C. for 1 h or until complete as determined by TLC or LC-MSanalysis. Solution was then diluted with aqueous, saturated NaHCO₃ (50mL) and the layers were separated. The aqueous layer was extracted withCH₂Cl₂ (1×25 mL) and the combined organics were dried (Na₂SO₄), filteredand concentrated in vacuo. Crude material was used as is or purified bynormal phase flash chromatography.

Purification A (1-200 mg Scale)

Crude product was purified by preparative scale reverse-phase HPLC(Phenomenex Gemini C-18 column, 110 Å, 30×100 mm, flow rate: 20 mL/min,mobile phase A: 0.1% TFA/water, mobile phase B: 0.1% TFA/ACN, gradientelution from 0% B to 100% B, UV (254 nm) and MS detection). Fractionscontaining the desired product were combined and lyophilized to providethe trifluoroacetate salt of target compound as a white solid.

Purification B (200-2500 mg Scale).

[Varian L/L 4002-2 column (5×50 cm.; Microsorb 100-10 C-18), flowrate=50 ml/min; mobile phase A: 100% water, 0.1% TFA; mobile phase B:100% ACN, 0.1% TFA; gradient elution from 20% B to 95% B in 90 min.,detection 254 nm].

Synthesis of (2S,3R)-2-amino-3-(Fmoc-amino)-butyric acid methyl esterhydrochloride (6)

Step 1: (2S,3S)-3-hydroxy-2-(trityl-amino)-butyric acid methyl ester (2)

A suspension of H-allo-Thr-OMe.HCl (1, 45.8 g, 270 mmol) in anhydrousCH₂Cl₂ (225 ml) was ice cooled. Anhydrous NEt₃ (85 ml, 610 mmol) wasadded, followed by a solution of trityl chloride (79 g, 284 mmol) inanhydrous CH₂Cl₂ (225 ml) (dropwise over 30 min). The reaction mixturewas allowed to warm to room temperature and stirred overnight. Theresulting suspension was filtered and the white solid was washed withCH₂Cl₂ (2×450 ml). The filtrate was combined with the first CH₂Cl₂washing and was washed with sat. NaHCO₃ (225 ml) and brine (225 ml). Thesecond CH₂Cl₂ washing was used to back extract the aqueous extracts. Thecombined CH₂Cl₂ solutions were dried (Na₂SO₄), filtered, andconcentrated. Hexane (450 ml) was added and the resulting mixture wasconcentrated. The resulting solid was triturated with hexane (600 ml),filtered, and washed with hexane to provide 2 (102.5 g, 100% yield, 98%pure by NMR) as a white solid.

Step 2: (2S,3R)-3-azido-2-(trityl-amino)-butyric acid methyl ester (3)

A solution of 2 (98.4 g, 262 mmol) and PPh₃ (68.7 g, 262 mmol) inanhydrous CH₂Cl₂ (1.7 l) was ice cooled. Diisopropyl azodicarboxylate(DIAD, 78 ml, 396 mmol) was added dropwise over 20 min, followed bydiphenylphosphoric azide (DPPA, 93 ml, 432 mmol) added dropwise over 20min. The resulting solution was allowed to warm to room temperature andstirred for two days. Solvent was evaporated and the residue waspurified twice by flash chromatography (2 kg of SiO₂ each, eluting with30% CH₂Cl₂ in hexanes, containing 1% NEt₃) to provide 3 (46.7 g, 45%yield) as a clear oil.

Step 3A: (2S,3R)-3-amino-2-(trityl-amino)-butyric acid methyl ester (4A)

To a solution of 3 (10 g, 25 mmol) in EtOAc (200 ml) was added Pd/C (5%wt, 2.5 g) and the resulting mixture was stirred under an H₂ balloon atroom temperature for two days. The reaction mixture was filtered throughCelite and the filtrate was concentrated to provide the desired compound4A (9.2 g, 99% yield) which was used directly in the next step.

Step 4A: (2S,3R)-3-(Fmoc-amino)-2-(trityl-amino)-butyric acid methylester (5A)

A solution of 4A (0.94 g, 2.5 mmol) and Fmoc-OSu (0.89 g, 2.63 mmol) inanhydrous THF (5 ml) was stirred at room temperature for 3.5 h. Solventwas evaporated and the resulting residue was purified by flashchromatography (CombiFlash, 40 g silica gel column, 35 ml/min, 10-30%EtOAc in hexane) to provide 5A (1.2 g, 81% yield) as a white solid.

Step 5A: (2S,3R)-2-amino-3-(Fmoc-amino)-butyric acid methyl ester (6)

To a solution of 5A (1.2 g, 2 mmol) in anhydrous THF (6 ml) was added 2M HO/ether (3 ml, 6 mmol) and the resulting solution was stirred at roomtemperature. A precipitate slowly begun forming. Anhydrous ether (9 ml)was added after 3.5 h and the resulting mixture was stirred at roomtemperature overnight. The solid was collected by filtration and waswashed with ether (15 ml) to provide 6 (0.7 g, ca. 90% yield). NMRshowed that this material contains about 4% of trityl alcohol.Trituration in ether is in progress.

Synthesis of (2S,3R)-2-amino-3-(Boc-amino)-butyric acid methyl esterhydrochloride (6A-HCl)

Note: Steps 1, 2 and 3A were described in the experimentals for (6)above.

Step 4B: (2S,3R)-3-(Boc-amino)-2-(trityl-amino)-butyric acid methylester (5B)

A solution of 4A (137 g, 366 mmol) and Boc anhydride (96 g, 438 mmol) inanhydrous CH₂Cl₂ (350 ml) was stirred at room temperature overnight.Solvent was evaporated and the resulting residue was purified by flashchromatography (SiO₂, 10-30% EtOAc/hexane) to provide 5B (111.8 g, 64%yield) as a white solid. ¹H-NMR (300 MHz, DMSO-d₆) δ 7.42 (bd, J=7.2 Hz,6H), 7.26 (bt, J=7.2 Hz, 6H), 7.18 (bt, J=7.2 Hz, 3H), 6.98 (bd, J=8.8Hz, 1H), 3.9-3.75 (m, 1H), 3.24 (dd, J=11.3, 5.8 Hz, 1H), 3.01 (s, 3H),2.66 (d, J=11.3 Hz, 1H), 1.37 (s, 9H), 1.09 (d, J=6.9 Hz, 3H).

Step 5B: (2S,3R)-2-amino-3-(Boc-amino)-butyric acid methyl ester (6A)

To a solution of 5B (126.8 g, 267 mmol) in anhydrous MeOH (1.4 l) wasadded Pd/C (10%, wet, 19.5 g) and the resulting mixture was hydrogenatedin a Parr apparatus (50 psi pressure) over two days. The reactionmixture was filtered through Celite and solvent was evaporated. Theresidue was purified by flash chromatography (SiO₂, 50-100%EtOAc/hexane, then 0-10% MeOH/EtOAc) to provide 6A (57.6 g, 92% yield)as an oil. ¹H-NMR (300 MHz, DMSO-d₆) δ 6.62 (bd, J=9.1 Hz, 1H), 3.9-3.75(m, 1H), 3.57 (s, 3H), 3.28 (d, J=4.1 Hz, 1H), 1.69 (bs, 2H), 1.36 (s,9H), 1.03 (d, J=6.9 Hz, 3H)

Step 5C: (2S,3R)-2-amino-3-(Boc-amino)-butyric acid methyl ester HClsalt (6A-HCl)

A solution of 6A (57.6 g, 248 mmol) in anhydrous ether (300 ml) was icecooled. A solution of HCl in ether (2 M, 124 ml, 248 mmol) was addeddropwise over 15 minutes. The resulting solution was stirred for 5minutes, hexane (900 ml) was added, and the resulting solid wascollected by filtration. The solid was washed with hexane and was driedunder high vacuum to provide 6A-HCl (63.3 g, 95% yield) as a whitesolid. ¹H-NMR (300 MHz, DMSO-d₆) δ 8.53 (bs, 3H), 6.96 (d, J=8.3 Hz,1H), 4.15-4.05 (m, 2H), 3.70 (s, 3H), 1.38 (s, 9H), 1.13 (d, J=6.9 Hz,3H). MS (APCI, pos): 233 (M+1, 20%), 177 (233-CH₂═CMe₂, 100%), 133(177-CO₂). Elemental Analysis: Found: C, 44.16; H, 7.90; N, 10.06; Cl,13.03. Calc.: C, 44.10; H, 7.92; N, 10.29; Cl, 13.02(C₁₀H₂₁N₂O₄Cl.0.2H₂O). LC purity: 100% (MS, TIC).

(2S, 3S)-methyl3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-2-aminobutanoatehydrochloride (7) (2R,3S)-methyl3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-2-aminobutanoatehydrochloride (8) (2R,3R)-methyl3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-2-aminobutanoatehydrochloride (9)

(7), (8) and (9) were prepared using the same method as described in thesynthesis of compound (6).

Synthesis of 2-amino-3-(Boc-amino)-3-methyl-butyric acid methyl ester(C001)

Synthesis of 2-amino-3-nitro-3-methyl-butyric acid methyl ester (1)

-   Reference: J. Chem. Soc. Perkin Trans. 1, 2659-2660 (1999).

2-Nitropropane (4.1 mL, 46 mmol, 2 eq) was taken up in 60 mL watercontaining KOH (3 g, 54 mmol, 2.3 eq). A solution of glyoxylic acidhydrate (2.15 g, 23 mmol, 1 eq) was treated with 30 mL of ammonia water(28-30%, ca. 10 eq), and this was added to the mixture containing thenitropropane. The clear, colorless reaction mixture was stirredvigorously for three hours at room temperature. The reaction mixture wascooled in ice water and treated with conc. HCl (aq) to pH=1-2. Thereaction mixture attained a blue color at ca. pH 4 but remainedhomogeneous. The blue color was removed by washing sequentially with2×50 mL and 1×25 mL chloroform, then the aqueous layer was stripped toyield a white solid. The solid was then taken up in one liter ofabsolute ethanol. Insoluble material was removed by filtration anddiscarded, and the filtrate was stripped to give2-amino-3-methyl-3-nitrobutanoic acid (1) as a white solid, which wasused without further purification.

This material was taken up in methanol and cooled in ice. HCl gas wasbubbled through for ca. one hour, and the reaction mixture was stirredat room temperature overnight. A white solid formed, which was removedby filtration and deemed not to be product by ¹H NMR. The methanol wasremoved in vacuo and the resulting slightly greasy off-white solid takenup in 750 mL of chloroform and stirred with 500 mL of sat. NaHCO₃ (aq.)to remove HCl. The layers were separated and the chloroform layer driedover Na₂SO₄, then stripped to an oil. Pure product was obtained viachromatography on silica gel using 5% ethyl acetate/hexanes. Afterremoval of solvent, the resulting yellow oil was dried under vacuum forno more than one hour to avoid loss of material. No solvents weredetected in the ¹H NMR. Yield: 24 g (50%). TLC Rf=0.31 (40% ethylacetate/hexanes). APCI(+) m/z=177, 130 amu (the latter peak is the mostprominent, and is due to loss of HNO₂ occurring in the massspectrometer). ¹H NMR (CDCl₃, 300 MHz) δ 4.15 (s, 1H), 3.74 (s, 3H),1.59 (s, 1H), 1.58 (s, 1H). C₆H₁₂N₂O₄ requires C, 40.91; H, 6.87; N,15.90. Found C, 41.50; H, 6.99; N, 15.34.

2-(Z-amino)-3-methyl-3-nitro-butyric acid methyl ester (2)

To a solution of 2-Amino-3-nitro-3-methyl-butyric acid methyl ester 1(5.37 g, 30.5 mmol) in 60 mL of THF and DIEA (6.6 mL, 40 mmol) Z—OSu(7.23 g, 29 mmol) was added and the reaction mixture was stirred 4 h atr.t. THF was removed under reduced pressure, the residue was taken inEtOAc (200 mL). Organics was washed with water (50 mL×2), brine (50 mL)and dried over Na₂SO₄. The solvent was removed by rotary evaporation andcolorless oil 2 (5.9 g, 62%), which was used on next step withoutpurification.

2-(Z-amino)-3-amino-3-methyl-butyric acid methyl ester (3)

To a cold (ice bath) solution of compound 2 (5.9 g, 19 mmol) in 10 mL ofAcOH and 40 ml of THF, Zn dust (16.5 g, 250 mmol, 13 eq) was added over2 min. After adding of zinc dust, the temperature of the reactionmixture was allowed to rise to ambient. Reaction was monitored by LCMS.After 2-3 hrs, LCMS showed the major peak of amine along with a smallpeak of acid (˜5%). The reaction mixture was filtered through Celiteusing THF to rinse, and the filtrate was concentrated by rotaryevaporation to give thick syrup. (Note: this syrup contained some amountof Zn(OAc)₂, but it would not affect next reaction).

2-(Z-amino)-3-(Boc-amino)-3-methyl-butyric acid methyl ester (4)

The above syrup was re-dissolved in 40 ml THF. Boc₂O (9.1 g, 41.8 mmol,2.2 eq) was added, followed by addition of DIEA (10 ml, 57 mmol, 3 eq).The reaction mixture was stirred for 3 hr at ambient temperature. WhenLC-MS indicated the completion of reaction, solvent was evaporated andthe resulting residue was purified by flash chromatography (Combiflash,80 g Si gel column, 40 ml/min, 0-30% EtOAc in Hexanes) to provide thedesired compound (5.3 g, 74% on two steps) as white crystal.

2-Amino-3-(Boc-amino)-3-methyl-butyric acid methyl ester (C001)

To a solution of compound 4 (5.2 g, 14 mmol) in 70 ml of MeOH was addedPd/C (5% wt, 800 mg) and the resulting mixture was hydrogenated using aParr shaker (50 psi H₂) for 30 min. Catalyst was removed by filtrationand the filtrate was concentrated to give target compound C001 (3.3 g,as white crystal, 97.3%).

(2S)-2-Amino-3-(tert-butoxycarbonylamino)-3-methyl-butyric acid methylester HCl salt Via S-alpha-Methylbenzylamine (E-(S)—HCl)

Synthesis of 3-Methyl-3-nitro-(2(S)-(1(S)-phenylethyl-amino))-butyricacid (A-(S))

The 2-nitropropane (90.87 g, 1.02 mol, 1.02 eq) and water (1.0 L) wereplaced under nitrogen in a 2 L Erlenmeyer flask with a large stirbar(2″×⅜″). With good stirring, the potassium hydroxide (˜90%; 74.81 g,1.20 mol, 1.2 eq) was added all at once (solution warms to ˜40° C.). Theflask was placed in a regulated water or oil bath heated to 45° C. The(S)-alpha-methylbenzylamine (130 mL, 1.02 mol, 1.02 eq) was measured andadded very quickly (very fast CO₂ absorption). The reaction mixture wasmaintained at ˜44-46° C. and stirred swiftly as the glyoxylic acid (50%aq, 112 mL, 1.00 mol, 1.00 eq) was added slowly dropwise (60 min,slowest for the last one-third) via a dropping funnel. The reactionmixture becomes cloudy, then clear, and when the solids begin formingagain the addition was slowed down. After complete addition, thereaction was stirred for an additional 2-4 h under nitrogen as thetemperature cools to 25-30° C. Additional water (200 mL) was added andthe reaction mixture warmed to 34° C. (internal) and stirred swiftly asthe 3.00 M aq hydrochloric acid (610 mL, 1.83 mol, 1.83 eq) was warmedin a dropping funnel and added as a stream over 20-30 min. The thickslightly off-white suspension was stirred for 0-16 h at room temperature(and if necessary, then placed in a −10° C. bath to cool to ˜18-20° C.internally). The cooled suspension was then filtered with suctionthrough paper and rinsed with dilute aq hydrochloric acid (0.2 M, 2 L),water (2 L) and ethyl ether (500 mL). The filtercake was then suctionedto compact “dryness” over 30-60 min. The filtercake was then transferredto a 20° C. desiccator or vacuum oven and dried under full vacuum for2-6 h. The solids were then ground and placed in the vacuum oven for 2-4h at 50-60° C., and 2-4 h at 90° C. to yield A-(S) as a slightlyoff-white powder (127.0 g, 47.7% yield). Check of a sample by HPLC-MS*showed a 4.5:95.5 ratio of diastereomers (2.13 min (minor), 2.30 min(major)) of the product with an overall purity of 94%. Both peaks showthe same molecular weight (MH⁺=266.7). LC-MS [M+H] 267.7 (C13H18N2O4+H,requires 267.30). *[Common HPLC method: Onyx monolithic C18 column,50×4.6 mm; 1.5 mL/min; 9.10 min gradient of 5%-60% MeCN in H₂O with 0.1%TFA; UV (254 nm); MS; ELSD. Retention times are reported for UV (254 nm)unless otherwise noted.]

Purification of 3-Methyl-3-nitro-(2(S)-(1(S)-phenylethyl-amino))-butyricacid (A-(S))

A solution of dilute aq hydrochloric acid (3.0 M, 250 mL), water (3.0 L)and acetic acid (350 mL) was placed in a 4 L Erlenmeyer flask andstirred well as it was immersed in a 45-60° C. bath and warmed to 40° C.(internal). Once the solution was up to temperature, dissolve theamino-acid A-(S) (˜10:90 d.r., 33.03 g) in stirred warm DMSO (50° C.,dry, 150 mL) and add 100 mL of acetic acid to form a clear solution.Pour the solution into a separatory (dropping) funnel that was warmedand contains 150 mL of warm acetic acid and 100 mL of warm DMSO and mix.Suspend the dropping funnel over a fast vortex in the Erlenmeyer flaskand add the warm DMSO solution to the Erlenmeyer flask at an evendropwise rate (˜5-10 mL/min, ˜45-60 min addition). The suspension wasthen filtered through paper by suction and rinsed with dilute aqhydrochloric acid (0.2 M, 2 L), water (4 L), isopropanol (150 mL) andethyl ether (500 mL). The filtercake was then suctioned to compact“dryness” over 20-40 min. The filtercake was then transferred to a 20°C. vacuum oven and dried under full vacuum for 3-12 h. The solids arethen ground and placed in the vacuum oven for 2 h at 50-60° C., and 2-4h at 90-95° C. to yield A-(S) as a pale tan-white powder (29.66 g, 89.8%yield). Check of a sample by HPLC-MS showed a 1.2:98.8 ratio ofdiastereomers (HPLC-MS: 2.12 min (minor), 2.29 min (major); bothMH⁺=267.6) of the product with an overall purity >99%.

Second Purification of3-Methyl-3-nitro-(2(S)-(1(S)-phenylethyl-amino))-butyric acid (A-(S))

(Procedure was similar to the first precipitation above.) A solution ofdilute aq hydrochloric acid (3.0 M, 250 mL), water (3.0 L) and aceticacid (450 mL) was placed in a 4 L Erlenmeyer flask and stirred well asit was immersed in a 45-60° C. bath and warmed to 40° C. (internal).Once the solution was up to temperature, dissolve the amino-acid A-(S)(1.2:98.8 d.r., 31.05 g) in stirred warm DMSO (50° C., dry, 175 mL) andadd 100 mL of acetic acid to form a clear solution. Pour the solutioninto a separatory (dropping) funnel that was warmed and contains 100 mLof warm acetic acid and 75 mL of warm DMSO and mix. Suspend the droppingfunnel over a fast vortex in the Erlenmeyer flask and add the warm DMSOsolution to the Erlenmeyer flask at an even dropwise rate (˜5-10 mL/min,˜45-60 min addition). Upon complete addition, the suspension was stirredand placed in a −10° C. bath to cool to ˜20-22° C. internally. Thesuspension was then filtered through paper by suction and rinsed withdilute aq hydrochloric acid (0.2 M, 2 L), water (4 L), isopropanol (150mL) and ethyl ether (500 mL). The filtercake was then suctioned tocompact “dryness” over 20-40 min. The filtercake was then transferred toa 20° C. vacuum oven and dried under full vacuum for 3-12 h. The solidsare then ground and placed in the vacuum oven for 2 h at 50-60° C., and2-4 h at 90-95° C. to yield A-(S) as a slightly off-white powder (29.6g, 95% yield). Check of a sample by HPLC-MS showed a 0.15:99.85 ratio ofdiastereomers (2.12 min (minor), 2.31 min (major)) of the product withan overall purity >99%.

Synthesis of 3-Methyl-3-nitro-2(S)-(1(S)-phenylethyl-amino)-butyric acidmethyl ester (B—(S))

Into a dry 100 mL flask with a stirbar was placed the amino-acid A-(S)(9.77 g, 36.7 mmol) and cesium carbonate (12.55 g, 38.05 mmol, 1.05 eq.)under nitrogen. With rapid stirring the dimethylformamide (37 mL) wasadded rapidly and stirred for 10 min, with sonication for 1 min. Thereaction mixture was then cooled in a 10-15° C. water bath as the firstportion of iodomethane (1.75 mL, two-thirds of the 2.63 mL) was addeddropwise over 15 min. The bath was removed and the reaction was broughtto room temperature over 20 min. The second portion of iodomethane (0.88mL, one-third of the 2.63 mL) was added dropwise more slowly over 30min. After the reaction was stirred for an additional 20 min, check of asample by HPLC-MS showed a major product peak (5.89 min, MH⁺=281.7;99.2% convn) with of a small amount of the SM (2.64 min, ˜0.75%). Afterthe reaction was stirred for an additional 60 min, check of a sample byHPLC-MS showed complete conversion to the product peak. The reaction wasthen washed with EtOAc and water into a separatory funnel containingEtOAc (250 mL), water (70 mL) and 3 M aq HCl (12.8 mL, 38.4 mmol). Thephases were shaken and the aqueous layer was adjusted to pH˜7-8 andseparated. The organic phase was washed with 3% aq Li₂SO₄ (3×100 mL),half-saturated aq NaHCO₃ (50 mL) and satd aq NaCl (2×100 mL), dried(Na₂SO₄), filtered and evaporated under reduced pressure with a heptanechaser to an amber oil. After placing under full vacuum for >4 h, theamino-ester B—(S) (9.88 g, 96% yield) was obtained. Check of a sample byHPLC-MS showed a major product peak (3.32 min, MH⁺=281.7). LC-MS [M+H]281.7 (C14H20N2O4+H, requires 281.33).

Synthesis of 3-Amino-3-methyl-(2(5)-(1(S)-phenylethyl-amino))-butyricacid methyl ester (C—(S))

The amino-ester B—(S) (9.88 g, 35.24 mmol) was dissolved in dry THF (100mL) and glacial acetic acid (150 mL) along with activated powderedmolecular sieves (4 A, 12 g) and stirred mildly for 2-4 h undernitrogen. The flask was then immersed in a 0° C. bath and stirred wellfor 20 min. To the cooled reaction mixture was then added the zinc dust(20.74 g, 317 mmol, 9 eq) portionwise: 1 eq of zinc dust (2.30 g) wasadded and the flask removed from the bath and stirred for 10 min, andthen immersed in a 20° C. bath, stirred for 20 min. Another 2 eq portionof zinc dust (4.61 g) was added and the mixture stirred ˜20-30 min untilslight venting through a bubbler ceased. The last sequence was repeated3 more times to finish the addition of zinc dust (9 eq total). Afterstirring a total of 18 h, a check of a sample by HPLC-MS showed a majorproduct peak (3.36 min, MH⁺=251.7) with a minor diamino-acid peak (1.38min, MH⁺=237.6). This mixture was then diluted with THF (˜150 mL) andfiltered through a celite pad with additional THF (˜300 mL) washing.This slightly cloudy solution was rotary evaporated under full vacuum toyield a slightly yellow oily solid (32.6 g). This material was dissolvedin 3:1 chloroform/isopropanol (200 mL) and poured into a separatoryfunnel containing 3:1 chloroform/isopropanol (500 mL) and 0.25 MEDTAsolution at pH 10.5-11 (275 mL). Aqueous 4 MNaOH soln (18 mL) was addedand the mixture was shaken and the pH of the aqueous checked. Additional4 M NaOH soln was added in portions to reach pH 10.5-11.0. The funnelcontents were thoroughly shaken, and the aqueous phase separated. Theorganic phase was then thoroughly shaken with 0.25 MEDTA solution at pH10.5-11 (1×200 mL, 1×50 mL), and separated. The organic phase was thenwashed with satd aq NaCl (3×150 mL), dried (Na₂SO₄), filtered andevaporated under reduced pressure, followed with addition of heptane(3×100 mL) and evaporation to yield a light-amber oil. After placingunder full vacuum for ˜1-2 h, the diamino-ester C—(S) (7.62 g, 86.4%yield) was obtained. Check of a sample by HPLC-MS showed a major productpeak (3.32 min, MH⁺=251.7). LC-MS [M+H] 251.7 (C14H22N2O2+H, requires251.34).

Synthesis of 3-(tert-Butoxycarbonylamino)-3-methyl-(2(S)-(1(S)-phenylethyl- amino))-butyric acid methyl ester (D-(S))

The diamino-ester C—(S) (7.62 g, 30.44 mmol, 1.0 eq) was dissolved inTHF (150 mL) under nitrogen and diisopropylethylamine (5.8 mL, 33.5mmol, 1.10 eq) was added to this solution. The tert-butylpyrocarbonate(9.96 g, 45.66 mmol, 1.50 eq) was added in two portions: the firstportion (7.97 g, 1.20 eq) was followed by the second portion (1.99 g,0.30 eq) 1 h later. After stirring for 10 h, a check of a sample byHPLC-MS showed a major product peak (4.94 min, MH⁺=351.7) with a minorSM peak (3.59 min, ˜2%). After stirring for 16 h, a check of a sample byHPLC-MS showed complete reaction. The reaction mixture was thendissolved in EtOAc (˜300 mL) and washed with water containing 1 eq HCl(0.3 M HCl, 100 mL), half saturated NaHCO₃ soln (100 mL), ˜14% aq NH₄OH(2×75 mL), satd aq NaCl (2×70 mL), dried (Na₂SO₄), filtered andconcentrated under reduced pressure to yield the BOC-diamino-ester D-(S)(10.5 g) as an oily part crystalline residue. This material wasdissolved in a minimum of DCM and loaded onto a Combiflash column(silica gel, 230-400 mesh, 400 g) and chromatographed using DCM (85mL/min) as eluant for 10 min, followed with a gradient of EtOAc (0-10%over 40 min) in DCM. The pure fractions eluted between 0.6% and 5% EtOAcin DCM. Evaporation gave 6.30 g of pure BOC-diamino-ester D-(S) as aslightly crystalline viscous oil. Check of a sample by HPLC-MS showed amajor product peak (4.85 min, MH⁺=351.7). LC-MS [M+H] 351.7(C19H30N2O4+H, requires 351.46).

Synthesis of (2S)-2-Amino-3-(tert-butoxycarbonylamino)-3-methyl-butyricacid methyl ester (E-(S))

The combined BOC-diamino-ester D-(S) (6.30 g, 17.98 mmol) was dissolvedin THF (5% soln, 125 mL) in Parr shaker bottle and placed undernitrogen. The palladium hydroxide catalyst (2.27 g, 18 wt %) was rapidlyweighed and added to the shaker flask with a following flush withnitrogen. The shaker flask was then connected to hydrogen and evacuatedand refilled 3 times. The flask was then filled with hydrogen (70 psi)and shaking was commenced. The pressure was kept above 60 psi as thehydrogen was consumed. After 1 h, check of a sample by HPLC-MS showedincomplete deprotection. After 11 h, check of a sample by HPLC-MS showeda major product peak (2.55 min (MS-TIC), 2.59 (ELSD), MH⁺=247.5) with noother peaks visible by UV (254 nm), MS, or ELSD. This mixture was thencarefully flushed with nitrogen and filtered through a celite pad withadditional MeOH (˜250 mL) washing. This clear solution was rotaryevaporated under reduced pressure (to 35° C.) followed with addition ofheptane (50 mL) and evaporation to yield a “water” clear oil (4.2 g)that was kept under vacuum. LC-MS [M+H] 247.5 (C11H22N2O4+H, requires247.31).

Synthesis of (2S)-2-Amino-3-(tert-butoxycarbonylamino)-3-methyl-butyricacid methyl ester HCl salt (E-(S)—HCl)

The BOC-Me₂DAP methyl ester (E-(S)) (506 mg, 2.05 mmol) under nitrogenwas dissolved in diethyl ether (3 mL), cooled in a 0° C. bath andrapidly stirred as a 2 M in Et₂O hydrochloric acid soln (1.05 mL, 2.10mmol, 1.02 eq) was added over 1 min. After stirring an additional 4 min,hexanes (25 mL) was added, and the suspension shaken, centrifuged anddecanted. Additional hexanes (˜20 mL) was added and the suspensionshaken, cooled in a 0° C. bath, centrifuged and decanted. The whiteprecipitate was dried under full vacuum to yield the BOC-diamino-esterhydrochloride salt (E-(S)—HCl) (592 mg, 101.9% yield). Check of thesample by HPLC-MS showed the major product peak (2.89 min (MS-TIC), 2.92(ELSD), MH⁺=247.5) with no other peaks visible by UV (254 nm), MS, orELSD. LC-MS [M+H] 247.5 (C11H22N2O4+H, requires 247.31).

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* E-(S)-HCl 2.05 592 100 100 247.5 2.89 *[HPLC method: Onyxmonolithic C18 column, 50 × 4.6 mm; 1.5 mL/min; 9.10 min gradient of5%-60% MeCN in H₂O with 0.1% TFA; UV (254 nm); MS; ELSD. Retention timesreported for UV (254 nm).]

Synthesis of (2R)-2-Amino-3-(Boc-amino)-3-methyl-butyric acid methylester (C002)

(C002) was prepared using the same method described in the synthesis ofcompound (E-(S)—HCl) by R-alpha-Methylbenzylamine instead.

Synthesis of di-Me-(S)-Ser methyl ester

Thionyl chloride (0.34 ml, 4.7 mmol) was added dropwise to a suspensionof H—(S)-di-Me-Ser-OH (250 mg, 1.88 mmol) in MeOH (4 ml) at −10° C.Reaction mixture was stirred at low temperature for 30 min and thentemperature of reaction was raised to ambient. After the reactionmixture was heated at 50° C. for 2 days. Then, additional amount ofthionyl chloride (0.34 ml, 4.7 mmol) was added to the reaction mixtureat low temperature. Stirring was continued at 50° C. for 3 days.Completion of the reaction was monitored by TLC (in order to check astarting amino acid) and LCMS. After completion (5-6 days) solvent wasevaporated in vacuum and residue was dried in vacuum overnight toprovide target compound hydrochloride (343 mg, 100%) as white solid.Compound was used as is for the next step transformation with noadditional purification.

Example 82N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(4-methoxy-phenyl)-buta-1,3-diynyl]-benzamide(82-1)

Synthesis of 1-ethoxymethyl-4-ethynyl-benzene (1a)

To a cold solution of 4-ethynyl benzyl alcohol (500 mg, 3.8 mmol) andiodoethane (336 mL, 4.2 mmol) in anhydrous THF (3 mL) at 0° C. was addedNaH (212 mg, 5.3 mmol). The reaction mixture was stirred at 25° C. for30-60 min, monitored by LC-MS until there was no starting materialremained. The solution was quenched with MeOH (1 mL) and volatiles wereremoved in vacuo. The crude mixture was partitioned in aqueous HCl (1 N,10 mL) and EtOAc (10 mL), the aqueous layer was extracted with ethylacetate and the combined organics were washed with brine, dried(Na₂SO₄), filtered and concentrated in vacuo. Crude material waspurified by flash chromatography (20 g SiO₂, EtOAc/Hex) to give thedesired compound (300 mg, 61%) as light yellow oil.

Synthesis of 1-(Fmoc-amino)-4-ethynyl-benzene (1b)

To a cold solution of 4-ethynyl aniline (1.4 g, 11.9 mmol) and DIEA (33mL, 20 mmol) in CH₂Cl₂ (20 mL) at 0° C. was added Fmoc-Cl (3.25 g, 5.3mmol) over 2 min. The solution was stirred at ambient temperatureovernight. Solution was concentrated in vacuo and the crude material wasredissolved in EtOAc (200 mL) and washed with water (2×70 mL) and brine(1×100 mL), dried (Na₂SO₄), filtered and concentrated in vacuo toprovide the desired compound as a brown solid (3.35 g, 83%).[M+H]⁺=439.1; Ret. Time (Method A)=6.78 min.

Synthesis of 4-[4-(4-Methoxy-phenyl)-buta-1,3-diynyl]-benzoic acidmethyl ester (2)

Prepared as described in Method 1-B using 1c (2.0 g, 15.1 mmol) andmethyl 4-(2,2-dibromovinyl)benzoate (4.00 g, 12.5 mmol). Purified bynormal phase flash chromatography (80 g SiO₂, 0-50% EtOAc/Hex) toprovide the desired compound 2 (940 mg, 26%) as an off-white solid.LC-MS: [M+H]⁺=Not observed; Ret. Time (Method A) 7.76 min; ¹H NMR (250MHz, CDCl₃) δ 3.8 (3H, s), 3.9 (3H, s), 6.8 (2H, d), 7.45 (2H, d), 7.6(2H, d), 8.0 (2H, d).

Synthesis of 4-[4-(4-Methoxy-phenyl)-buta-1,3-diynyl]-benzoic acid (3)

Prepared as described in Method 2-A using compound 2 (470 mg, 1.62 mmol)to provide the desired compound 3 (412 mg, 92%) as an off-white solid.LC-MS: [M+H]⁺=Not observed; Ret. Time (Method A) 6.34 min.

Synthesis of (2S,3R)-3-tert-Butoxycarbonylamino-2-{4-[4-(4-methoxy-phenyl)-buta-1,3-diynyl]-benzoylamino}-butyric acidmethyl ester (4)

Prepared as described in Method 3-A using compound 3 (150 mg, 0.543mmol) and (2S,3R)-3-methyl Dap.HCl (160 mg, 0.598 mmol) to provide thedesired compound 4 (448 mg, 168%) as a viscous amber oil. LC-MS:[M+H]⁺=491.2; Ret. Time (Method A) 6.99 min.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(4-methoxy-phenyl)-buta-1,3-diynyl]-benzamide(82-1)

Prepared as described in Method 5-A using compound 4 (266 mg, 0.543mmol) to provide the crude product (119 mg, 56%) as an off-white solid.Crude product was purified by Purification Method A to provide the TFAsalt of desired compound 82-1 (9 mg, 3%) as a white solid. LC-MS:[M+H]⁺=392.4; Ret. Time (Method A) 4.34 min.

Scale Yield Yield Purity [M + Retention Compound (mmol)¹ (mg) (%)¹ (%)²H] time (min)² 82-1 0.543 8.5 3.1 99.1 392.4 4.34 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.Each of the following compounds was synthesized as described above usingthe appropriate phenyl acetylene (all commercially available with theexception of 1a and 1b), amino acid and deprotection.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 82-2 

4.79 378.3 A 82-3 

4.95 392.3 A 82-4 

6.67 362.3 A 82-5 

6.87 380.3 A 82-6 

7.78 430.3 A 82-7 

6.58 380.3 A 82-8 

7.47 395.9 A 82-9 

7.36 395.9 A 82-10

6.97 396.3 A 82-11

7.90 446.3 A 82-12

6.79 380.3 A 82-13

6.48 377.1 A 82-14

5.60 393.1 A 82-15

5.89 407.1 A 82-16

4.10 377.1 A 82-17

4.90 392.3 A 82-18

7.55 381.1 A 82-19

7.78 395.1 A 82-20

7.71 421.1 A 82-21

8.03 435.1 A 82-22

5.70 431.1 A 82-23

5.13 381.1 A 82-24

A 82-25

A 82-26

A 82-27

7.05 394.3 A 82-28

7.01 394.3 A 82-29

6.83 394.3 A

Example 83N—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-[4-(4-hydroxy-phenyl)-buta-1,3-diynyl]-benzamide(83-1)

Synthesis of (4-tert-butoxy-phenylethynyl)-trimethyl-silane (2)

A 20 mL microwave tube was charged with a mixture of1-bromo-4-tert-butoxybenzene 1 (1.0 g, 4.36 mmol), PdCl₂(dppf) (71 mg,0.087 mmol), CuI (33 mg, 0.174 mmol), DIPA (0.924 mL, 6.55 mmol), andTHF (15 mL). Tube was backfilled with nitrogen, sealed, and irradiatedin a microwave reactor (max. power 250 W) at 100° C. for 5 min. Solutionwas diluted with water (200 mL) and extracted with EtOAc (3×80 mL).Combined organic layers were washed with water (1×100 mL) and brine(1×80 mL), dried (Na₂SO₄), filtered and concentrated in vacuo. Crudeproduct was purified by normal phase flash chromatography (40 g SiO₂,0-30% EtOAc/Hex) to provide the desired compound 2 (488 mg, 45%) as aclear colorless oil. LC-MS: [M+H]⁺=Not observed; Ret. Time (Method A)7.87 min; ¹H NMR (250 MHz, CDCl₃) δ 0.2 (s, 9H), 1.3 (s, 9H), 6.9 (d,2H), 7.4 (d, 2H).

Synthesis of 1-tert-Butoxy-4-ethynyl-benzene (3)

Prepared as described in Method 3-A using compound 2 (488 mg, 1.98 mmol)to provide the desired compound 3 (373 mg, 108%) as a clear colorlessoil. LC-MS: [M+H]⁺=Not observed; Ret. Time (Method A) 5.87 min; ¹H NMR(250 MHz, CDCl₃) δ 1.3 (s, 9H), 3.0 (s, 1H), 6.9 (d, 2H), 7.4 (d, 2H).

Synthesis of 4-[4-(4-t-Butoxy-phenyl)-buta-1,3-diynyl]-benzoic acidmethyl ester (4)

Prepared as described in Method 1-B using compound 3 (373 mg, 2.14 mmol)and methyl 4-(2,2-dibromovinyl)benzoate (822 mg, 2.57 mmol) to providethe desired compound 4 (160 mg, 23%) as an off-white solid. LC-MS:[M+H]⁺=Not observed; Ret. Time (Method A) 8.28 min; ¹H NMR (250 MHz,CDCl₃) δ 1.35 (s, 9H), 3.9 (s, 3H), 6.9 (d, 2H), 7.45 (d, 2H), 7.6 (d,2H), 8.0 (d, 2H).

Synthesis of 4-[4-(4-tert-Butoxy-phenyl)-buta-1,3-diynyl]-benzoic acid(5)

Prepared as described in Method 2-A using compound 4 (160 mg, 0.482mmol) to provide the desired compound 5 (130 mg, 85%) as a slightlyoff-white solid. LC-MS: [M+H]⁺=Not observed; Ret. Time (Method A) 7.10min.

Synthesis of (S)-3-tert-Butoxycarbonylamino-2-{4-[4-(4-tert-butoxy-phenyl)-buta-1,3-diynyl]-benzoylamino}-3-methyl-butyricacid methyl ester (6)

Prepared as described in Method 3-A using compound 5 (65 mg, 0.205 mmol)and (2S,3R)-3-methyl Dap (56 mg, 0.227 mmol) to provide the desiredcompound 6 (117 mg, 104%) as a light amber solid. LC-MS: [M+H]⁺=547.2;Ret. Time (Method A) 8.23 min.

Synthesis ofN—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-[4-(4-hydroxy-phenyl)-buta-1,3-diynyl]-benzamide(83-1)

Prepared as described in Method 5-A using compound 6 (112 mg, 0.205mmol) to provide the desired compound 83-1. Crude material was purifiedby Purification Method A to provide the TFA salt of the desired compound83-1 (36 mg, 35%) as a white solid. LC-MS: [M+H]⁺=391.8; Ret. Time(Method A) 3.65 min.

Scale Yield Yield Purity [M + Retention Compound (mmol)¹ (mg) (%)¹ (%)²H] time (min)² 83-1 0.205 36.2 34.9 99.2 391.8 3.65 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.The following compound was synthesized as described above.

Compound # Structure RT¹ (min) [M + H] 83-2

3.53 378.4 ¹Using LC-MS Analytical Method A.

Example 84N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(4-cyclopropoxy-phenyl)-buta-1,3-diynyl]-benzamide(84-1)

Synthesis of 1-Bromo-4-(2-chloro-ethoxy)-benzene (1)

To a mixture of 4-bromophenol (2.0 g, 11.6 mmol) and K₂CO₃ (3.20 g, 23.2mmol) in DMF (50 mL) was added 3-chloroethyl p-toluenesulfonate (2.52mL, 13.9 mmol) and the mixture was stirred at 60° C. for 16 h. Reactionmixture was concentrated in vacuo, and the residue was partitionedbetween water (200 mL) and CH₂Cl₂ (50 mL). Aqueous layer was furtherextracted with CH₂Cl₂ (3×20 mL). Combined organic layers were washedwith water (30 mL) and brine (30 mL), dried (Na₂SO₄), filtered andconcentrated in vacuo to provide the desired compound 1 (3.12 g, 115%)as a light amber oil. LC-MS: [M+H⁺]=Not observed; Ret. Time (Method A)5.83 min; ¹H NMR (250 MHz, CDCl₃) δ 3.8 (t, 2H), 4.2 (t, 2H), 6.8 (d,2H), 7.4 (d, 2H).

Synthesis of 1-Bromo-4-vinyloxy-benzene (2)

To a stirred solution of compound 1 (2.7 g, 11.6 mmol) in THF (30 mL),at 0° C., was added potassium tert-butoxide (1.95 g, 17.3 mmol) in threeportions. After 10 min the solution was allowed to warm to ambienttemperature and stirred for 16 h. The solution was then concentrated invacuo and partitioned between water (150 mL) and CH₂Cl₂ (50 mL). Aqueouslayer was further extracted with CH₂Cl₂ (3×30 mL). Combined organiclayers were washed with water (40 mL) and brine (40 mL), dried (Na₂SO₄),filtered and concentrated in vacuo. Crude material was purified bynormal phase flash chromatography (40 g SiO₂, 100% Hex) to provide thedesired compound 2 (1.70 g, 74%) as a clear oil. LC-MS: [M+H]⁺=Notobserved; Ret. Time (Method A) 5.95 min; ¹H NMR (250 MHz, CDCl₃) δ 4.45(dd, 1H), 4.75 (dd, 1H), 6.6 (dd, 1H), 6.85 (d, 2H), 7.4 (d, 2H).

Synthesis of 1-Bromo-4-cyclopropoxy-benzene (3)

To a stirred solution of compound 2 (1.70 g, 8.55 mmol) and CH₂I₂ (5.52mL, 68.4 mmol) in anhydrous 1,2-dichloroethane (75 mL), under N₂ at 0°C. was cautiously added a solution of diethylzinc (1.0 M, 68.4 mL) inhexanes. After 30 min the solution was allowed to warm to ambienttemperature and stirred for 64 h. Solution was then added cautiously tosaturated, aqueous NH₄Cl (200 mL). The mixture was shaken and layersseparated, and aqueous layer was extracted with CH₂Cl₂ (3×30 mL).Combined organic layers were washed with aqueous HCl (1.0 N, 1×100 mL),water (1×100 mL), saturated NaHCO₃ (aq) (1×100 mL) and brine (1×80 mL),dried (Na₂SO₄), filtered and concentrated in vacuo to. Crude product waspurified by normal phase flash chromatography (40 g SiO₂, 100% Hex) toprovide the desired compound 3 (1.31 g, 72%) as a clear oil. LC-MS:[M+H]⁺=Not observed; Ret. Time (Method A) 5.87 min; ¹H NMR (250 MHz,CDCl₃) δ 0.75 (m, 4H), 3.7 (m, 1H), 6.9 (d, 2H), 7.4 (d, 2H).

Synthesis of (4-Cyclopropoxy-phenylethynyl)-trimethyl-silane (4)

To a mixture of compound 3 (900 mg, 4.22 mmol), PdCl₂(Ph₃P)₂ (150 mg,0.211 mmol), CuI (80 mg, 0.420 mmol), Ph₃P (1.44 g, 5.49 mmol), and DIPA(5 mL, 35.4 mmol) in DMF (10 mL), in a 20 mL microwave tube, was addedethynyltrimethylsilane (0.716 mL, 5.07 mmol). Tube was backfilled withN₂, sealed, and irradiated in a microwave reactor (max. power 250 W) at120° C. for 15 min. Solution was diluted with water (200 mL) andextracted with EtOAc (3×60 mL). Combined organic layers were washed withwater (1×100 mL) and brine (1×60 mL), dried (Na₂SO₄), filtered andconcentrated in vacuo. Crude product was purified by normal phase flashchromatography (40 g silica gel, 0-10% EtOAc/Hex) to provide a mixtureof target compound 4 (840 mg) and triphenylphosphine. LC-MS: [M+H]⁺=Notobserved; Ret. Time (Method A) 7.52 min.

Synthesis of 1-Cyclopropoxy-4-ethynyl-benzene (5)

To a solution of compound 4 (972 mg, 4.22 mmol) in MeOH (15 mL) wasadded NaOH (400 mg, 10 mmol) and the mixture was stirred at ambienttemperature for 20 min. Solution was diluted with water (150 mL), cooledto 0° C., neutralized with aqueous HCl (1.0 N), and extracted withCH₂Cl₂ (4×40 mL). Combined organic layers were washed with water (1×80mL) and brine (1×80 mL), dried (Na₂SO₄), filtered and concentrated invacuo. Crude material was purified by preparative scale reverse-phaseHPLC (Varian Microsorb 100-10 C-18 column (50×300 mm), flow rate: 50mL/min, mobile phase A: 0.1% TFA/water, mobile phase B: 0.1% TFA/ACN,gradient elution from 10% B to 80% B over 90 min, UV 254 nm monitor).Fractions containing the desired product were combined and neutralizedwith NaHCO₃ (aq). Acetonitrile was removed by evaporation in vacuo, andthe residual aqueous solution was extracted with EtOAc (3×40 mL). Thecombined organic layers were washed with water (1×60 mL) and brine (1×60mL), dried (Na₂SO₄), filtered and concentrated in vacuo to provide thedesired compound 5 (34 mg, 5%) as a clear oil. LC-MS: [M+H]⁺=Notobserved; Ret. Time (Method A) 5.55 min; ¹H NMR (250 MHz, CDCl₃) δ 0.75(m, 4H), 3.0 (s, 1H), 3.7 (m, 1H), 7.0 (d, 2H), 7.4 (d, 2H).

Synthesis of 4-[4-(4-Cyclopropoxy-phenyl)-buta-1,3-diynyl]-benzoic acidmethyl ester (6)

Prepared as described in Method 1-B using compound 5 (34 mg, 0.216 mmol)and methyl 4-(2,2-dibromovinyl)benzoate (83 mg, 0.259 mmol) to providethe desired compound 6 (40 mg, 59%) as an off-white solid. LC-MS:[M+H]⁺=Not observed; Ret. Time (Method A) 8.03 min.

Synthesis of 4-[4-(4-Cyclopropoxy-phenyl)-buta-1,3-diynyl]-benzoic acid(7)

Prepared as described in Method 2-A using compound 6 (40 mg, 0.127 mmol)to provide the desired compound 7 (33 mg, 86%) as an off-white solid.LC-MS: [M+H]⁺=Not observed; Ret. Time (Method A) 6.81 min.

Synthesis of (2S,3R)-3-tert-Butoxycarbonylamino-2-{4-[4-(4-cyclopropoxy-phenyl)-buta-1,3-diynyl]-benzoylamino}-butyricacid methyl ester (8)

Prepared as described in Method 3-A using compound 7 (33 mg, 0.109 mmol)and (2S,3R)-3-methyl Dap (32 mg, 0.120 mmol) to provide the desiredcompound 8 (89 mg, 158%) as a viscous amber oil. LC-MS: [M+H]⁺=517.4;Ret. Time (Method A) 7.55 min.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(4-cyclopropoxy-phenyl)-buta-1,3-diynyl]-benzamide(84-1)

Prepared as described in Method 4-A followed by Method 5-A usingcompound 8 (56 mg, 0.109 mmol) to provide the desired compound 84-1.Crude material was purified by Purification Method A to provide the TFAsalt of the desired compound 84-1 (25 mg, 44%) as a white solid. LC-MS:[M+H]⁺=418.4; Ret. Time (Method A) 4.84 min.

Scale Yield Yield Purity [M + Retention Compound (mmol)¹ (mg) (%)¹ (%)²H] time (min)² 84-1 0.109 25.3 43.6 95.1 418.4 4.84 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.

Example 85 Synthesis ofN—[(S)-2-(2-Cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-(4-phenyl-buta-1,3-diynyl)-benzamide(85-1)

Synthesis of 4-(4-Phenyl-buta-1,3-diynyl)-benzoic acid methyl ester (1)

Prepared as described in Method 1-B using phenylacetylene (760 mg, 7.44mmol) and methyl 4-(2,2-dibromovinyl)benzoate (1.70 g, 5.31 mmol) toprovide the desired compound 1 (1.14 g, 82%) as an off-white solid.

Synthesis of 4-(4-Phenyl-buta-1,3-diynyl)-benzoic acid (2)

Prepared as described in Method 2-A using compound 1 (1.14 g, 4.37 mmol)to provide the desired compound 2 (995 mg, 92%) as a white solid.

Synthesis of(S)-3-tert-Butoxycarbonylamino-2-[4-(4-phenyl-buta-1,3-diynyl)-benzoylamino]-propionic acid methyl ester (3)

Prepared as described in Method 3-A using compound 2 (100 mg, 0.40 mmol)and (2S) (Boc)Dap.HCl (111 mg, 0.42 mmol) to provide the desiredcompound 3 (127 mg, 70%) as a white solid. LC-MS: [M+H]⁺=447.3; Ret.Time (Method A) 6.87 min.

Synthesis of(S)-3-Amino-2-[4-(4-phenyl-buta-1,3-diynyl)-benzoylamino]-propionic acidmethyl ester (4)

Prepared as described in Method 4-A using compound 3 (127 mg, 0.28 mmol)to provide the desired compound 4 (100 mg, 95%). LC-MS: [M+H]⁺=347.4;Ret. Time (Method A) 4.82 min.

Synthesis of(S)-3-[2-(tert-Butoxycarbonyl-cyclopropyl-amino)-acetylamino]-2-[4-(4-phenyl-buta-1,3-diynyl)-benzoylamino]-propionicacid methyl ester (5)

Prepared as described in Method 3-A using compound 4 (100 mg, 0.40 mmol)and N-cyclopropyl-N-Boc-glycine.HCl (111 mg, 0.42 mmol) to provide thedesired compound 3 (127 mg, 70%). LC-MS: Ret. Time (Method A) 5.34 min.

Synthesis of (S)-3-(2-Cyclopropylamino-acetylamino)-2-[4-(4-phenyl-buta-1,3-diynyl)-benzoylamino]-propionicacid methyl ester (6)

Prepared as described in Method 4-A using compound 5 (127 mg, 0.23 mmol)to provide the desired compound 6. LC-MS: [M+H]⁺=444.5; Ret. Time(Method A) 4.99 min.

Synthesis ofN—[(S)-2-(2-Cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-(4-phenyl-buta-1,3-diynyl)-benzamide(85-1)

Prepared as described in Method 5-B using compound 6 (235 mg, 0.40 mmol)to provide the desired compound 85-1. Crude material was purified byPurification Method A to provide the TFA salt of the desired compound85-1 (16 mg, 7% yield) as a white solid. LC-MS: [M+H]⁺=445.1; Ret. Time(Method A) 6.75 min.

Retention Scale Yield Yield Purity [M + time Compound (mmol)¹ (mg) (%)¹(%)² H] (min)² 85-1 0.4 15.5 6.9 98.4 445.1 6.75 ¹Based on the amount ofcarboxylic acid used in the coupling reaction. ²Using LC-MS AnalyticalMethod A.The following compound was synthesized as described above using theappropriate amino acid and deprotection.

HPLC- MH⁺ Ret. Time MS Compound # Structure (m/z) (min) Method 85-2

459.1 6.84 A

Example 86 Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-{4-[(2,2-difluoro-ethylamino)-methyl]-phenyl}-buta-1,3-diynyl)-benzamide(86-1)

Synthesis of(2S,3R)-3-tert-Butoxycarbonylamino-2-{4-[4-(4-hydroxymethyl-phenyl)-buta-1,3-diynyl]-benzoylamino}-butyricacid methyl ester (2)

Prepared as described in Method 3-A using compound 1 (390 mg, 1.4 mmol)and (S)-Boc-Me-Dap-OMe.HCl (403 mg, 1.5 mmol) to provide the desiredcompound 2 (660 mg, 96%). LC-MS: [M+H]⁺=491.2; Ret. Time (Method A) 5.98min.

Synthesis of(2S,3R)-3-tert-Butoxycarbonylamino-2-[4-(4-{4-[(2,2-difluoro-ethylamino)-methyl]-henyl}-buta-1,3-diynyl)-benzoylamino]-butyricacid methyl ester (4)

To a solution of compound 2 (660 mg, 1.35 mmol) and TEA (0.42 mL, 3.0mmol) in CH₂Cl₂ (6 mL) at 0° C., methanesulfonyl chloride (113 mg, 1.45mmol) was added. The solution was then allowed to warm to ambienttemperature and stirred for 1 h. To this solution containing compound 3was added 2,2-difluoroethylamine (273 mg, 3.37 mmol) dropwise andstirred overnight at 50° C. All volatiles were removed in vacuo and thecrude material was diluted with EtOAc (100 mL) and washed with water(2×40 mL), brine (1×40 mL), dried (Na₂SO₄), filtered and concentrated invacuo to provide the desired compound 4. LC-MS: [M+H]⁺=554.5; Ret. Time(Method A) 4.81 min.

Synthesis of (2S,3R)-3-Amino-2-[4-(4-{4-[(2,2-difluoro-ethylamino)-methyl]-phenyl}-buta-1,3-diynyl)-benzoylamino]-butyricacid methyl ester (5)

Prepared as described in Method 4-A using compound 4 (374 mg, 0.68 mmol)to provide the desired compound 5. LC-MS: [M+H]⁺=454.8; Ret. Time(Method A) 3.07 min.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-{4-[(2,2-difluoro-ethylamino)-methyl]-phenyl}-buta-1,3-diynyl)-benzamide (86-1)

Prepared as described in Method 5-A using compound 5 (355 mg, 0.675mmol) to provide the desired compound 86-1. Crude material was purifiedby Purification Method A to provide the TFA salt of the desired compound86-1 (76 mg, 16%) as a white solid. LC-MS: [M+H]⁺=455.8; Ret. Time(Method A) 3.79 min.

Retention Scale Yield Yield Purity [M + time Compound (mmol)¹ (mg) (%)¹(%)² H] (min)² 86-1 0.7 75.7 18 98.6 455.8 3.79 ¹Based on the amount ofcarboxylic acid used in the coupling reaction. ²Using LC-MS AnalyticalMethod A.Each of the following compounds was synthesized as described above usingthe appropriate amine and amino acid.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 86-2

3.71 461.1 A 86-3

4.01 431.5 A 86-4

3.38 475.9 A 86-5

3.75 437.1 A 86-6

4.22 509.1 A 86-7

4.42 523.5 A

Example 87 Synthesis ofN-(2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-[4-(4-cyclopropylaminomethyl-phenyl)-buta-1,3-diynyl]-benzamide(87-1)

Synthesis of Cyclopropyl-(4-trimethylsilanylethynyl-benzyl)-amine (2)

To a solution of compound 1 (10 g, 48.9 mmol) and DIEA (16 mL, 96.5mmol) in CH₂Cl₂ (25 mL) at 0° C., was added methanesulfonyl chloride(4.56 mL, 58.7 mmol) dropwise. The solution was allowed to warm toambient temperature and stirred for 1 h. Cyclopropyl amine (25.69 g, 450mmol) was added dropwise and resulting solution was stirred overnight atambient temperature. The solution was concentrated in vacuo and thecrude material was diluted with EtOAc (200 mL), washed with water (2×70mL), brine (1×100 mL) and dried (Na₂SO₄), filtered and concentrated invacuo. The crude material was purified by normal phase chromatography(120 g SiO₂, CHCl₃/MeOH) to provide the desired compound 2 (3.6 g, 30%)as brown liquid. LC-MS: [M+H]⁺=244.2; Ret. Time (Method A) 4.86 min.

Synthesis of Cyclopropyl-(4-trimethylsilanylethynyl-benzyl)-carbamicacid tert-butyl ester (3)

To a solution of compound 2 (3.6 g, 14.8 mmol) and Et₃N (4.17 mL, 30mmol) in CH₂Cl₂ (15 mL), was added Boc₂O (3.55 g, 16.26 mmol) and theresulting mixture was stirred for 2 h at ambient temperature. Thesolution was concentrated in vacuo and the crude material was dilutedwith EtOAc (150 mL), washed with water (2×70 mL), brine (1×80 mL) anddried (Na₂SO₄), filtered and concentrated in vacuo to provide thedesired compound 3 (4.32 g) as a brown oil. LC-MS: [M+H]⁺=Not observed;Ret. Time (Method A) 8.31 min.

Synthesis of Cyclopropyl-(4-ethynyl-benzyl)-carbamic acid tert-butylester (4)

To a solution of NaOH (0.20 g, 5.0 mmol) in MeOH (75 mL), was addedcompound 3 (4.32 g, 12.6 mmol) and resulting mixture was stirred for 1 hat ambient temperature. The solution was concentrated in vacuo. and thecrude material was diluted with EtOAc (120 mL), washed with water (2×30mL), brine (1×50 mL), dried (Na₂SO₄), filtered and concentrated invacuo. The crude material was purified by normal phase flashchromatography (40 g SiO₂, EtOAc/Hex) to provide the desired compound 4(1.58 g, 47%) as white solid. LC-MS: [M+H]⁺=Not observed; Ret. Time(Method A) 6.38 min.

Synthesis of4-(4-{4-[(tert-Butoxycarbonyl-cyclopropyl-amino)-methyl]-phenyl}-buta-1,3-diynyl)-benzoicacid methyl ester (5)

Prepared as described in Method 1-B using 4 (1.07 g, 3.94 mmol) andmethyl 4-(2,2-dibromovinyl)benzoate (1.38 g, 4.33 mmol) to provide thedesired compound 5 (615 mg, 36%) as a white solid. LC-MS: [M+H]⁺=430.4;Ret. Time (Method A) 8.73 min.

Synthesis of 4-(4-{4-[(tert-Butoxycarbonyl-cyclopropyl-amino)-methyl]-phenyl}-buta-1,3-diynyl)-benzoicacid (6)

Prepared as described in Method 2-A using compound 5 (615 mg, 1.4 mmol)to provide the desired compound 6 (509 mg, 88%) as a white solid. LC-MS:[M+H]⁺=416.5; Ret. Time (Method A) 7.44 min.

Synthesis of 3-tert-Butoxycarbonylamino-2-[4-(4-{4-[(tert-butoxycarbonyl-cyclopropyl-amino)-methyl]-phenyl}-buta-1,3-diynyl)-benzoylamino]-3-methyl-butyricacid methyl ester (7)

Prepared as described in Method 3-A using compound 6 (400 mg, 0.96 mmol)and (25)-3-dimethylDap-(OMe) (249 mg, 1.01 mmol) to provide the desiredcompound 6 (585 mg, 96%) as a white solid. LC-MS: [M+H]⁺=644.5; Ret.Time (Method A) 8.47 min.

Synthesis of3-Amino-2-{4-[4-(4-cyclopropylaminomethyl-phenyl)-buta-1,3-diynyl]-benzoylamino}-3-methyl-butyricacid methyl ester (8)

Prepared as described in Method 4-A using compound 7 (585 mg, 0.91 mmol)to provide the desired compound 8 (407 mg). LC-MS: [M+H]⁺=444.5; Ret.Time (Method A) 3.42 min.

Synthesis ofN-(2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-[4-(4-cyclopropylaminomethyl-phenyl)-buta-1,3-diynyl]-benzamide(87-1)

Prepared as described in Method 5-A using compound 8 (407 mg, 0.91 mmol)to provide the crude compound 87-1. Crude product was purified byPurification Method A to provide the TFA salt of desired compound 87-1(266 mg, 18%) as a white solid. LC-MS: [M+H]⁺=445.5; Ret. Time (MethodA) 3.10 min.

Retention Scale Yield Yield Purity [M + time Compound (mmol)¹ (mg) (%)¹(%)² H] (min)² 87-1 0.96 266 53.3 99.6 445.1 3.10 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.Each of the following compounds was synthesized as described above usingthe appropriate amino acid.

Ret. HPLC- Compound [MH⁺] Time Ms # Structure (m/z) (min) Method 87-2

432.3 4.64 A 87-3

446.3 4.90 A 87-4

4.07 445.5 A

Example 88 Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(3-methyl-3H-imidazol-4-yl)-buta-1,3-diynyl]-benzamide(88-1)

Synthesis of 4-[4-(3-Methyl-3H-imidazol-4-yl)-buta-1,3-diynyl]-benzoicacid methyl ester (2)

Prepared as described in Method 1-B using5-ethynyl-1-methyl-1H-imidazole (220 mg, 2.10 mmol) and methyl4-(2,2-dibromovinyl)benzoate (700 mg, 2.18 mmol) and purified byPurification Method B to provide the TFA salt of the desired compound 2(482 mg, 59%) as a yellow solid. LC-MS: [M+H]⁺=265.1; Ret. Time (MethodA) 3.97 min.

Synthesis of 4-[4-(3-Methyl-3H-imidazol-4-yl)-buta-1,3-diynyl]-benzoicacid (3)

Prepared as described in Method 2-A (Method 2-B for pyridyl derivatives)using compound 2 (482 mg, 1.83 mmol) to provide the desired compound 3(334 mg, 73%) as an off-white solid. LC-MS: [M+H]⁺=250.9; Ret. Time(Method A) 3.13 min.

Synthesis of(2S,3R)-3-(9H-Fluoren-9-ylmethoxycarbonylamino)-2-{4-[4-(3-methyl-3H-imidazol-4-yl)-buta-1,3-diynyl]-benzoylamino}-butyricacid methyl ester (4)

Prepared as described in Method 3-A using compound 3 (100 mg, 0.40 mmol)and (2S,3R)Fmoc-3-Me-Dap(OMe) HCl (164 mg, 0.67 mmol) to provide thedesired compound 4 (235 mg). LC-MS: [M+H]⁺=586.7; Ret. Time (Method A)5.21 min.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(3-methyl-3H-imidazol-4-yl)-buta-1,3-diynyl]-benzamide(88-1)

Reagent MW, d Eq. mmol mg, ml Compound 4 586.65 1.0 0.4 235 mg NH2OH ×HCl 69.49 6.14 1.72 120 mg 25% MeONa in MeOH 0.5 mL THF 1 mL MeOH 2.5 mL

The Compound 88-1 was made using the General Method for hydroxamateformation. Check of a sample by HPLC-MS showed a major product peak(Ret. time (Method A) 2.03 min, MH⁺=366.2). Crude product was purifiedby preparative scale reverse-phase HPLC (Phenomenex Gemini C-18 column,110 Å, 30×100 mm, flow rate: 20 mL/min, mobile phase A: 0.1% TFA/water,mobile phase B: 0.1% TFA/ACN, gradient elution from 15% B to 40% B over60 min, MS detection). Fractions containing the desired product werecombined and lyophilized to provide the trifluoroacetate salt of targetcompound 88-1 (112.4 mg, 47.3% yield) as a white solid. LC-MS: RT(Method A) 2.13 min; [M+H] 366.3 (C₁₉H₁₉N₅O₃+H, requires 366.4).

Retention Scale Yield Yield Purity [M + time Compound (mmol)¹ (mg) (%)¹(%)² H] (min)² 88-1 0.4 112.4 47.3 98.8 366.2 2.13 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.

tert-Butyl thiazol-2-ylcarbamate 2

A solution of thiazol-2-amine (10.0 g, 0.10 mol, 1.0 equiv), (Boc₂)O(26.2 g, 0.12 mol, 1.2 equiv), and DIPEA (25.8 g, 0.20 mol, 2.0 equiv)in THF (150 mL) was stirred at 50° C. for 12 hours. The reaction mixturewas concentrated in vacuum and EtOAc (300 mL) was added. Then, theorganic layer was washed by water (250 mL×2) and brine (250 mL×2), dried(MgSO₄), filtrated, concentrated in vacuum to give target product (6.3g, 31.5%). [M+1]: 201.1

tert-Butyl 5-bromothiazol-2-ylcarbamate 3

A suspension of NBS (6.2 g, 34.6 mmol, 1.1 equiv) and tert-butylthiazol-2-ylcarbamate (6.3 g, 31.5 mmol, 1.0 equiv) in THF (100 mL) wasstirred at room temperature for 12 hours. Then, the reaction mixture wasfiltrated and purified by column chromatography to give target compound(6.5 g, 73.9%)

tert-Butyl 5-((trimethylsilyl)ethynyl)thiazol-2-ylcarbamate 4

A mixture of tert-butyl 5-bromothiazol-2-ylcarbamate (6.5 g, 23.3 mmol,1.0 equiv), ethynyltrimethylsilane (3.4 g, 34.9 mmol, 1.5 equiv),PdCl₂(PPh₃)₂ (0.16 g, 0.23 mmol, 0.01 equiv), CuI (0.14 mg, 0.84 mmol,0.03 equiv) in TEA (50 mL) was stirred at 80° C. for 4 hours undernitrogen. The reaction mixture was extracted with EtOAc (500 ml) andorganics was washed with water (200 mL×2) and brine (200 mL) Organiclayer was dried over Na₂SO₄ and evaporated. The residue was purified bycolumn chromatography to give target compound (1.5 g, 30%).

tert-Butyl 5-ethynylthiazol-2-ylcarbamate 5

To a solution of tert-butyl5-((trimethylsilyl)ethynyl)thiazol-2-ylcarbamate (1.50 g, 5.1 mmol, 1.0equiv) in THF (50 mL) was treated with KOH/methanol (10.2 mmol, 0.57g/10 mL, 2 equiv) slowly below 10° C., then the mixture was allowed toreact at ambient temperature for 1 hours. Then, the reaction mixtureconcentrated in vacuum. The residue was dissolved in EtOAc (100 mL). Theorganic layer was washed with water (50 mL×2) and brine (50 mL), driedover Na₂SO₄, filtered and concentrated in vacuum. The residue was driedin vacuum overnight to provide the title compound (1.1 g, 96%)

Methyl4-((2-(tert-butoxycarbonylamino)thiazol-5-yl)buta-1,3-diynyl)benzoate 6

A mixture of methyl 4-(2,2-dibromovinyl)benzoate (1.10 g, 5.4 mmol, 1.1equiv), tert-butyl 5-ethynylthiazol-2-ylcarbamate (1.70 g, 4.9 mmol, 1.0equiv), Pd₂(dba)₃ (9 mg, 0.01 mmol, 0.02 equiv), TMPP (14 mg, 0.02 mmol,0.04 equiv) and TEA (97 mg, 9.8 mmol, 2 equiv) in DMF (20 ml) wasstirred at 45° C. for 12 hours under nitrogen. The reaction mixture wasextracted with EtOAc (100 ml) and organics was washed with water (100ml×2) and brine (50 ml). Organic layer was dried over Na₂SO₄ andevaporated. The residue was purified by column chromatography to givetarget compound (0.9 g, 46%). ¹H-NMR: (400 MHz, DMSO-d₆): δ 1.50 (s,9H), 3.87 (s, 3H), 7.74 (dd, J=2.4 Hz, 6.4 Hz), 7.93 (s, 1H), 7.99 (dd,J=2.0 Hz, 6.0 Hz), 12.09 (s, 1H).

4-((2-(tert-Butoxycarbonylamino)thiazol-5-yl)buta-1,3-diynyl)benzoicacid 7

To a solution ofmethyl-4-((2-(tert-butoxycarbonylamino)thiazol-5-yl)buta-1,3-diynyl)benzoate (0.9 g, 2.4 mmol, 1 equiv) in THF (10 mL) and MeOH (10 mL), 2 MNaOH aq. (2.0 ml) was added and the reaction was stirred at roomtemperature for 1 h. After cooling the reaction mixture was acidified toa pH of 1-2 with 10% HCl. The formed solid was collected by filtration,washed with water (2×10 ml) and ice-cold MeOH (10 ml). The filtrationwas dried in vacuum overnight to give the final product (650 mg, 71%).[M+1]: 369.0; ¹H-NMR: (400 MHz, DMSO-d₆): δ 1.50 (s, 9H), 7.68 (d, J=6.0Hz), 7.91 (s, 1H), 7.97 (d, J=6.4 Hz). Compound 7 was used for synthesisof compound 88-17 and 88-18 using the appropriate amino acids.

Each of the following compounds was synthesized as described above usingthe appropriate amino acid and deprotection.

Ret. HPLC- Compound Time MH⁺ MS # Structure (min) (m/z) Method 88-2 

2.67 363.5 A 88-3 

3.44 363.5 A 88-4 

3.77 363.5 A 88-5 

4.30 378.3 A 88-6 

2.36 377.1 A 88-7 

3.41 378.3 A 88-8 

3.71 377.8 A 88-9 

2.84 367.1 A 88-10

3.13 381.1 A 88-11

 4.82¹ 383.3 A 88-12

 6.22² 367.9 B 88-13

 4.36² 382.0 B 88-14

4.77 383.2 A 88-15

4.31 382.0 B 88-16

4.95 383.1 A 88-17

3.08 383.9 A 88-18

3.08 383.9 A 88-19

401.4 88-20

405   expted 88-21

366.1 expted A 88-22

382   expted A 88-23

366.1 expted 88-24

382   expted

Example 89N-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((1-methyl-1H-pyrazol-4-yl)buta-1,3-diynyl)benzamide

Synthesis of 4-(2,2-dibromovinyl)-1-methyl-1H-pyrazole (1)

The 1-methyl-1H-pyrazole-4-carbaldehyde (1.00 g, 9.08 mmol) and carbontetrabromide (3.16 g, 9.54 mmol) were dissolved in CH₂Cl₂ (27 mL) andthe solution was cooled to 0° C. To this chilled solution was then addedPPh₃ (5.01 g, 19.1 mmol) in four portions over 5 min. The solution wasthen allowed to warm to ambient temperature for 18 h. The solution wasdiluted with hexanes (30 mL) and stirred for 15 min. The slurry wasfiltered and the filtrate was concentrated in vacuo and was purified byflash chromatography (50% EtOAc/Hex) to provide the desired product 1 asa white solid (1.72 g, 71%). LC-MS [M+H]⁺=266.8 (Theoretical, C₆H₇Br₂N₂⁺=266.9).

Synthesis of methyl4-((1-methyl-1H-pyrazol-4-yl)buta-1,3-diynyl)benzoate (2)

The methyl 4-ethynylbenzoate (1.09 g, 6.79 mmol) and compound 1 (1.72 g,6.47 mmol) were dissolved in DMF (65 mL) and this solution wasdeoxygenated by bubbling N₂ through for 10 min. To this deoxygenatedsolution was then added Et₃N (1.3 mL, 9.39 mmol). N₂ was then bubbledthrough for an additional 5 min followed by addition of Pd₂(dba)₃ (0.060g, 0.065 mmol) and tri(4-methoxyphenyl)phosphine (0.091 g, 0.259 mmol).The dark brown solution was then allowed to stir at 80° C. for 18 h thencooled to ambient temperature. The solution was diluted with EtOAc (100mL), washed with deionized water (3×50 mL) and brine (1×50 mL), dried(MgSO₄), filtered and concentrated in vacuo. The crude oil was purifiedby flash chromatography (0-50% EtOAc/CH₂Cl₂) to provide the desiredproduct 2 as a yellow solid (1.03 g, 60%). LC-MS [M+H]⁺=265.0(Theoretical, C₁₆H₁₃N₂O₂ ⁺=265.1).

Synthesis of 4-((1-methyl-1H-pyrazol-4-yl)buta-1,3-diynyl)benzoic acid(3)

To a solution of compound 2 (1.03 g, 3.90 mmol) in a mixture of THF (13mL), MeOH (13 mL), and water (13 mL) was added LiOH monohydrate (0.491g, 11.7 mmol) in one portion. The solution was allowed to stir atambient temperature for 18 h. The pH of the solution was adjusted withaqueous HCl (1.0 N) and aqueous NaHCO₃ (sat.) to pH˜7.5. The resultingsolution was then extracted with EtOAc (3×100 mL), dried (MgSO₄),filtered and concentrated in vacuo to provide the desired product 3(0.400 g, 41%) as an off white solid. LC-MS [M+H]⁺=251.0 (Theoretical,C₁₅H₁₁N₂O₂ ⁺=251.1).

Synthesis of (2S,3R)-methyl3-(tert-butoxycarbonylamino)-2-(4-((1-methyl-1H-pyrazol-4-yl)buta-1,3-diynyl)benzamido)butanoate(4)

Compound 3 (0.400 g, 1.60 mmol), (2S,3R)-methyl 3-(Boc)-2-aminobutanoateHCl (0.473 g, 1.76 mmol), EDC (0.613 g, 3.20 mmol) and HOBT (0.432 g,3.20 mmol) were slurried in anhydrous CH₂Cl₂ (16 mL). To this slurry wasadded N,N-diisopropylethylamine (1.04 mL, 6.40 mmol) in one portion. Thesolution was then allowed to stir at ambient temperature for 18 h. Thesolution was then concentrated in vacuo and the crude solid was purifiedby flash chromatography (0-50% EtOAc/CH₂Cl₂) to provide the desiredproduct 4 (0.785 g, 81%) as an off white solid. ¹H NMR (DMSO) δ 8.43 (d,J=8.4, 1H), 8.22 (s, 1H), 7.91 (d, J=7.9, 2H) 7.80 (s, 1H), 7.72 (d,J=7.3, 2H) 7.00 (d, J=9.7, 1H), 4.62 (dd, J=4.2, 8.2, 1H) 4.22-4.27 (m,1H) 3.86 (s, 3H), 3.61 (s, 3H), 1.36 (s, 9H), 1.11 (d, J=7.0, 3H).

Synthesis of (2S,3R)-methyl3-amino-2-(4-((1-methyl-1H-pyrazol-4-yl)buta-1,3-diynyl)benzamido)butanoate(5)

To a solution of compound 4 (0.286 g, 0.616 mmol) in of CH₂Cl₂ (10 mL)was added TFA (2 mL) slowly. The solution was allowed to stir at ambienttemperature for 1 h then concentrated in vacuo. The thick oil was thenazeotroped with CH₂Cl₂ (3×) to provide the TFA salt of the desiredproduct 5 (0.284 g, 100%) as an off white solid. LC-MS [M+H]⁺=365.1(Theoretical, C₂₀H₂₁N₄O₃ ⁺=365.2).

Synthesis ofN-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((1-methyl-1H-pyrazol-4-yl)buta-1,3-diynyl)benzamide(89-1)

The TFA salt of 5 (0.284 g, 0.616 mmol) was dissolved in isopropylalcohol (6 mL) and the hydroxylamine solution (0.81 mL, 12.3 mmol) wasadded in one portion at ambient temperature and the solution was allowedto stir for 18 h at ambient temperature. The volatiles were removed invacuo and the resulting crude material was purified by reverse phaseHPLC (5-30% MeCN/H₂O, w/0.1% TFA) then lyophilized to dryness to providethe desired product (89-1) (0.090 g, 40%) as a white solid; LC-MS[M+H]⁺=366.1 (Theoretical, C₁₉H₂₀N₅O₃ ⁺=366.2).

Each of the following compounds was synthesized as described above.

Ret. HPLC- Time MH⁺ MS Compound # Structure (min) (m/z) Method 89-2

4.71 380.3 A 89-3

5.43 381.1 A 89-4

3.99 364.3 A 89-5

2.46 380.3 A 89-6

4.26 378.3 A 89-7

4.96 379.1 A

Example 90 Synthesis of(S)-4-((1H-pyrazol-4-yl)buta-1,3-diynyl)-N-(3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)benzamide(90-1)

Synthesis of tert-butyl 4-iodo-1H-pyrazole-1-carboxylate (1)

NaH (247 mg, 6.18 mmol) was added to the DMF (50 ml) solution of4-iodo-1H-pyrazole (1.0 g, 5.15 mmol) at 0° C. The reaction was stirredat 0° C. in 30 min. (Boc)₂O (1.68 g, 7.76 mmol) was added to thereaction at 0° C. The reaction was warmed to r.t. nd stirred overnight.H₂O (200 ml) was added to quench the reaction. The mixture was extractedwith EtOAc (3×200 ml). The combined EtOAc extracts was washed with H₂O(2×200 ml), brine (200 ml) and dried with Na₂SO₄ the crude product waspurified with ISCO normal phase column (0-20% EtOAc/DCM). Compound 20(1.3 g) was obtained in the yield of 86%.

Synthesis of tert-butyl4-((trimethylsilyl)ethynyl)-1H-pyrazole-1-carboxylate (2)

THF was degassed by passing through N₂ gas for 30 min. TEA (6.0 ml, 11.5mmol) was added to the THF solution (20 ml) of Compound 1 (1.69 g, 5.75mmol), Trimethylsilylethyne (845 mg, 8.63 mml), PdCl₂ (PPh₃)₂ (404 mg,0575 mmol) and CuI (109 mg, 0.575 mmol). The reaction was stirredovernight. The reaction was diluted with EtOAc and filtered throughcelite. The organic solution was washed with H₂O, brine and dried withNa₂SO₄. The crude product was purified with ISCO normal phase column Thecrude product was purified with ISCO normal phase column (0-2%EtOAc/DCM). Compound 21 (1.02 g, 67.2%) was obtained

Synthesis of tert-butyl 4-ethynyl-1H-pyrazole-1-carboxylate (3)

TBAF (5.2 ml, 1.0M in THF) was added to the THF solution (10 ml) ofCompound 2 (1.02 g, 3.46 mmol) at 0° C. The reaction was stirred at 0°C. for 10 min. The THF was removed and the residue was dissolved inEtOAc. The EtOAc solution was washed with H₂O, brine and dried withNa₂SO₂. The crude produce was purified with ISCO normal phase column.The crude product was purified with ISCO normal phase column (0-30%EtOAc/hexane). Compound 3 (362.3 mg, 54.5%) was obtained.

Synthesis of methyl 4-(bromoethynyl)benzoate (4)

NBS (13.4 g, 75 mmol) was added to the acetone solution (250 ml) of4-ethynyl-benzoic methyl ester (8 g, 50 mmol) and CF₃COOAg (850 mg, 5mmol). The reaction was stirred at r.t for 2 hrs. The reaction waspoured into ice-water (500 ml) and stirred for 30 min. After separation,the H₂O layer was extracted with EtOAc (300 ml×3). The combined EtOAclayers was washed with H₂O (2×200 ml) and brine (200 ml) and dried withNa₂SO₄. The crude product was purified with ISCO normal phase column(0-50% EtOAc/hexane). Compound 27 (12.0 g, 100%) was obtained.

Synthesis of methyl4-((1-methyl-1H-pyrazol-4-yl)buta-1,3-diynyl)benzoate (5)

THF was degassed by passing through N₂ gas for 30 min. TEA was added tothe THF solution (10 ml) of Compound 3 (362 mg, 1.89 mmol), Compound 4(666 mg, 2.27 mmol), PdCl₂ (PPh₂)₂ (133 mg, 0.189 mmol), PPh₃ (25 mg,0.095 mmol) and CuI (36 mg, 0.189 mmol). The reaction was stirred atr.t. for 30 min followed by the addition of CuI. The reaction was heatedto 45° C. overnight. The reaction was diluted with EtOAc and filteredthrough celite. The organic solution was washed with H₂O, brine anddried with Na₂SO₄. The crude product was purified with ISCO normal phasecolumn. The crude product was purified with ISCO normal phase column(0-50% EtOAc/hexane). Compound 28 (150 mg, 19.7%) was obtained

Synthesis of4-((1-(tert-butoxycarbonyl)-1H-pyrazol-4-yl)buta-1,3-diynyl)benzoic acid(6)

Compound 6 (120 mg, 94%) was made using the General Method for basichydrolysis used to the next step without further purification.

Synthesis of methyl3-(tert-butoxycarbonylamino)-2-(4-(imidazo[1,2-a]pyridin-6-ylbuta-1,3-diynyl)benzamido)-3-methylbutanoate(7)

Compound 7 (100 mg, 48%) was coupled to BocDAP-OMe using the GeneralMethod for HATU coupling used in the next step without purification.

Synthesis of (S)-methyl2-(4-((1H-pyrazol-4-yl)buta-1,3-diynyl)benzamido)-3-amino-3-methylbutanoate(8)

The Compound 8 was made using the General Method for Boc deprotectionand used on next step without purification. Compound 8 (65 mg, 100%)

Synthesis of(S)-4-((1H-pyrazol-4-yl)buta-1,3-diynyl)-N-(3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)benzamide(90-1)

The target product (33 mg, 50%, m+z=366.66) was prepared by followingGeneral Method 3-A for hydroxamate Method (Hydroxyamide formation,aqueous).

The following compounds were synthesized by using above procedure withcorresponding acid and amine.

Ret. HPLC- Compound Time MH⁺ MS # Structure (min) (m/z) Method 90-2

393.7 A 90-3

416.6 A

Example 91N—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-hexa-1,3-diynyl-benzamide(91-1)

Synthesis of 4-Hexa-1,3-diynyl-benzoic acid methyl ester (2)

To a solution of 4-(2,2-Dibromo-vinyl)-benzoic acid methyl ester (1)(1.5 g, 4.69 mmol), Pd₂dba₃ (45 mg, 1 mol %), TMPP (60 mg, 4 mol %), andTEA (1.94 mL, 14.1 mmol) in DMF (15 mL), cooled in a dryice/acetonitrile bath, was introduced 1-butyne (0.75 mL, 9.4 mmol) viacondensation on a −78° C. cold finger under a nitrogen atmosphere.Reaction vessel was sealed and heated at 80° C. for 16 h. Reactionmixture was diluted with water (200 mL) and extracted with EtOAc (3×80mL). Combined organic layers were washed with water (100 mL) and brine(80 mL), dried over anhydrous Na₂SO₄, and concentrated in vacuo to givea sticky brown solid. Crude product was purified by normal phase flashchromatography on a CombiFlash® Companion™ unit equipped with Luknovaflash column (40 g silica gel, 40-60 μm average particle size, 60 Å poresize); flow rate: 40 mL/min; mobile phase A: hexane; mobile phase B:EtOAc; gradient elution from 0% B to 30% B over 70 min. Fractionscontaining the desired product were combined and concentrated in vacuoto give target compound 2 (1.093 mg, 110%) as a slightly sticky lightbrown solid. LC-MS: RT (Method A) 6.67 min; compound not significantlyionizable. ¹H NMR (250 MHz, CDCl₃) δ 1.15 (3H, t), 2.35 (2H, q), 3.9(3H, s), 7.6 (2H, d), 8.0 (2H, d).

Synthesis of 4-Hexa-1,3-diynyl-benzoic acid (3)

To compound 2 (4.69 mmol) were added THF (8 mL) and 3 M NaOH (aq) (12mL, 36 mmol). Reaction mixture was heated at 80° C. for 1 h. Reactionmixture was diluted with water (150 mL), cooled in ice/water bath,acidified to pH 3 with 10% H₃PO₄ (aq), and filtered. Solids were washedwith water (3×50 mL) and dried by lyophilization to give target compound3 (766.9 mg, 83.4% from 1) as an off-white solid. LC-MS: RT (Method A)5.38 min; compound not significantly ionizable.

Synthesis of(S)-3-tert-Butoxycarbonylamino-2-(4-hexa-1,3-diynyl-benzoylamino)-3-methyl-butyricacid methyl ester (4)

Reagent MW Eq. mg/mL mmol Compound 3 198.22 1.0 96.1 mg 0.485 Chiraldimethyl DAP 246.31 1.1 133 mg 0.540 HATU 380.2 1.2 224 mg 0.589 DIEA129.25 3.0 0.256 mL 1.47 DMF 4 mL

Compound 4 (231.6 mg, 112%) as a sticky amber solid prepared using theGeneral Method for HATU coupling and used on next step withoutpurification. LC-MS: RT (Method A) 6.90 min; [M+H] 427.1 (C₂₄H₃₀H₂O₅+H,requires 427.53).

Synthesis ofN—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-hexa-1,3-diynyl-benzamide(91-1)

To compound 4 (˜0.485 mmol) was added 4 M HCl/dioxane (4 mL), and themixture was stirred at ambient temperature for 1 h. Volatiles wereremoved in vacuo to give a sticky amber solid. LC-MS: RT (Method A) 4.31min; [M+H] 327.1 (C₁₉H₂₂N₂O₃+H, requires 327.41). Isopropyl alcohol (4mL) was added to the solid and the mixture was cooled in an ice/waterbath for 5 min. NH₂OH (50%, aq) (4 mL) was added to the mixture,dropwise for the first 2 mL. Reaction mixture was allowed to stir in icebath for 5 min, and then allowed to stir at ambient temperature for 16h. Solvent volume was reduced approximately by half under a stream ofnitrogen, and water (10 mL) was added. The suspension was thoroughlyagitated (vibro mixer and sonication), centrifuged and the supernatantwas discarded. Water (10 mL) was added to the solid and the suspensionwas thoroughly agitated, centrifuged and the supernatant was discarded.Wet solid was dried by lyophilization to give crude product (127 mg,80.3% crude yield from 3) as an off-white solid. Crude product waspurified by preparative scale reverse-phase HPLC (Phenomenex Gemini C-18column, 110 Å, 30×100 mm, flow rate: 20 mL/min, mobile phase A: 0.1%TFA/water, mobile phase B: 0.1% TFA/ACN, gradient elution from 15% B to40% B over 60 min, MS detection). Fractions containing the desiredproduct were combined and lyophilized to provide the trifluoroacetatesalt of target compound 91-1 (64.8 mg, 30.3% yield from 3, 37.8%recovery from crude product) as a white solid. LC-MS: RT (Method A) 3.77min; [M+H] 328.3 (C₁₈H₂₁N₃O₃+H, requires 328.40).

Retention Scale Yield Yield Purity [M + time Compound (mmol)¹ (mg) (%)¹(%)² H] (min)² 91-1 0.485 64.8 30.3 98.5 328.3 3.77 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.

Intermediate Acid IC-1

Synthesis of cyclobutylacetylene (2)

The synthesis was done according to U.S. Pat. No. 6,303,057 entitled“Preparation of cycloalkylacetylene compounds”.

Into a 0.5 liter round bottom flask equipped with an addition funnel anda reflux condenser were placed), diisopropylamine (10.1 g, 0.1 mol) andTHF (250 ml). Bromoethane (59.9 g, 0.55 mol) was added dropwise to thismixture via the addition funnel at such a rate as to maintain arefluxing solution. Ethane was evolved during the addition. The reactionwas determined to be complete once all of the magnesium was consumed.This process produced a solution containing EtMgBr and (iPr)₂NMgBr.6-Chlorohexyne (23.32 g, 0.2 mol) was added dropwise and the resultingreaction mixture was stirred at 65° C. for 36 h. The cooled reactionmixture was poured onto crashed ice. Organic upper layer was separated,washed 3 times with water (3×100 mL) and dried over Na₂SO₄. Na₂SO₄ wasseparated by filtration to give 21.7 g of yellow liquid, which was usedon next step without purification.

Synthesis of 4-(4-Cyclobutyl-buta-1,3-diynyl)-benzoic acid (IC-1)

Reagent MW Eq. mmol g, ml Methyl 4-(2,2-dibromovinyl)- 319.98 1.0 7.82.5 g benzoate Cyclobutylacetylene (2) 80.13 1.4 10.93 0.88 gTris(dibenzyllidene- 915.72 0.08 0.065 60 mg acetone)dipalladium(0)Tris(4-methoxyphenyl)phosphine 352.36 0.03 0.24 85 mg TEA 101.19 3.2 253.47 mL DMF(anhydrous) 10 mL

The 4-(4-Cyclobutyl-buta-1,3-diynyl)-benzoic acid methyl ester (4) wasmade and separated using the General Method for Sonogashira coupling*.Yield: 11.3%. (Yellow solid). Check of a sample by HPLC-MS showed amajor product peak (Ret. time (Method A) 7.58 min, [MH+]=239.2). Thisproduct was subject to basic hydrolysis to give the desired acid IC-1,used to prepare compound 10-16 and 10-17.

For syntheses of the following alkyl- and cycloalkyl diacetylenes,commercially available acetylenes were used.

Ret. HPLC- Compound Time MH⁺ MS # Structure (min) (m/z) Method 91-2

5.46 326.3 A 91-3

6.79 354.3 A 91-4

7.35 368.3 A 91-5

6.12 327.1 A 91-6

6.50 341.1 A 91-7

3.64 314.1 A 91-8

4.04 328.3 A 91-9

4.14 342.2 AEach of the following compounds was synthesized as described above usingthe Boc protected amino acid and HCl deprotection.

Ret. HPLC- Time MH⁺ MS Compound # Structure (min) (m/z) Method 91-10

4.01 340.3 A 91-11

3.97 339.9 A 91-12

3.95 340.3 A 91-13

4.16 328.3 A 91-14

6.95 368.3 A 91-15

6.08 342.6 A 91-16

6.32 354.3 A 91-17

3.18 340.2 A

Intermediate Acid IC-2

Methyl 4-(7-methylocta-1,3-diynyl)benzoate (2)

To a degassed solution of compound 1 (2.8 g, 11 mmol), Pd(PPh₃)₂Cl₂(0.411 g, 0.58 mmol) and CuI (0.11 g, 0.58 mmol) in dried THF (50 mL)was added 5-methylhex-1-yne (1.35 g, 1.86 mL, 14 mmol), followed bydiisopropylamine (3.56 g, 4.9 ml, 35.1 mmol) dropwise. After theaddition of diisopropylamine, the reaction mixture was stirred overnightat room temperature. The suspension was filtered and washed the cakewith CH₂Cl₂. The filtrate was concentrated in vacuo. The residue waspurified by silica-gel column chromatography (PE: EA=100:1) to givecompound 2 (2.25 g, 75%) as a brown solid.

4-(7-Methylocta-1,3-diynyl)benzoic acid (3)

Reagent MW Eq. mmol g, mL Compound 2 254.32 1 8.6 2.15 g 2N NaOH 40 2.320 10 mL THF 30 mL MeOH 10 mL

Compound 3 (1.5 g, 70%) as a white solid made and separated using theGeneral Method for basic hydrolysis. ¹H-NMR (400 MHz, DMSO-d₆): 0.89 (d,J=6.4, 6H), 1.39-1.45 (m, 2H), 1.61-1.70 (m, 1H), 2.44-2.51 (m, 2H),7.65 (d, J=8.4 Hz, 2H), 7.93 (d, J=8.8 Hz, 2H), 13.24 (s, 1H).

The intermediate acids for making the following analogues weresynthesized by using the same synthetic method for compound 10-18. Eachof the following compounds of was synthesized as described above usingthe Boc protected amino acid and HCl deprotection.

Ret. HPLC- Compound Time MH⁺ MS # Structure (min) (m/z) Method 91-18

8.55 371.1 A 91-19

7.84 357.1 A 91-20

8.73 383.1 A 91-21

7.50 370.3 A 91-22

6.90 356.3 A 91-23

7.69 382.3 A 91-24

6.67 342.3 A 91-25

7.64 368.3 A 91-26

6.43 354.3 A 91-27

6.22 340.3 A 91-28

7.31 356.3 A 91-29

355.2 91-30

344.1

Example 92N-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((R)-5-morpholinohexa-1,3-diynyl)benzamideANDN-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((S)-5-morpholinohexa-1,3-diynyl)benzamide(92-1 and 92-2)

Synthesis of (rac)-methyl 4-(5-hydroxyhexa-1,3-diynyl)benzoate (1)

The methyl 4-(2,2-dibromovinyl)benzoate (3.00 g, 9.38 mmol) wasdissolved in DMF (50 mL) and this solution was deoxygenated by bubblingN₂ through for 10 min. To this deoxygenated solution was then added Et₃N(1.3 mL, 9.39 mmol) and 2-methyl-3-butyn-2-ol (0.342 g, 4.07 mmol). N₂was then bubbled through for an additional 5 min followed by addition ofPdCl₂(PPh₃)₂ (0.022 g, 0.031 mmol) and P(tBu)₃ HBF₄ (0.036 g, 0.125mmol). The dark brown solution was then allowed to stir at 85° C. for 3h then cooled to ambient temperature. The solution was diluted withEtOAc (100 mL), washed with deionized water (3×50 mL) and brine (1×50mL), dried (MgSO₄), filtered and concentrated in vacuo. The crude oilwas purified by flash chromatography (0-50% EtOAc/Hex) to provide thedesired product 1 as a brown solid (1.10 g, 71%). LC-MS [M+H]⁺=229.1(Theoretical, C₁₅H₁₃O₂ ⁺=229.1).

Synthesis of (rac)-methyl 4-(5-morpholinohexa-1,3-diynyl)benzoate (2)

Compound 1 (0.400 g, 1.75 mmol) was dissolved in anhydrous CH₂Cl₂ (20mL) under N₂ and cooled to 0° C. To this chilled solution was addedmethanesulfonic anhydride (0.336 g, 1.93 mmol) in one portion followedby slow addition of Et₃N (0.37 mL, 2.63 mmol). The solution was allowedto stir at 0° C. for 30 min then morpholine (5 mL) was added and thesolution was heated to 50° C. for 30 min. The solution was then removedfrom heat and allowed to stir at ambient temperature for 18 h. Thesolution was concentrated in vacuo and the crude oil was purified byflash chromatography (0-50% EtOAc/CH₂Cl₂) to provide the desired product2 (0.468 g, 90%) as a solid. LC-MS [M+H]⁺=298.1 (Theoretical, C₁₈H₂₀NO₃⁺=298.1).

Synthesis of (racy-4-(5-morpholinohexa-1,3-diynyl)benzoic acid (3)

To a solution of compound 2 (0.468 g, 1.57 mmol) in a mixture of THF (5mL), MeOH (5 mL), and water (5 mL) was added LiOH monohydrate (0.198 g,4.71 mmol) in one portion. The solution was allowed to stir at ambienttemperature for 4 h. The volatile organics were then carefully removedand the pH of the resulting aqueous solution was adjusted with NaHSO₄(1.0 N) to ˜2. The resulting slurry was vacuum filtered and the solidwas washed with deionized water (1×20 mL) to provide the desired product3 (0.445 g, 99%) as an offwhite solid. LC-MS [M+H]⁺=284.1 (Theoretical,C₁₇H₁₈NO₃ ⁺=284.1).

Synthesis of (2S,3R)-methyl3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-2-(4-((R)-5-morpholinohexa-1,3-diynyl)benzamido)butanoateand (2S,3R)-methyl3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-2-(4-((S)-5-morpholinohexa-1,3-diynyl)benzamido)butanoate (4)

Compound 3 (0.445 g, 1.57 mmol), (2S,3R)-methyl3-(Fmoc)-2-aminobutanoate (0.676 g, 1.73 mmol), EDC (0.602 g, 3.14 mmol)and HOBT (0.424 g, 3.14 mmol) were slurried in anhydrous CH₂Cl₂ (15 mL).To this slurry was added N,N-diisopropylethylamine (1.02 mL, 6.28 mmol)in one portion. The solution was then allowed to stir at ambienttemperature for 18 h. The solution was then diluted with water (20 mL)and CH₂Cl₂ (15 mL), then the layers were separated. The aqueous layerwas extracted with CH₂Cl₂ (3×10 mL) and the combined organics were dried(MgSO₄), filtered and concentrated in vacuo. The crude solid waspurified by flash chromatography (0-10% MeOH/CH₂Cl₂) to provide thedesired mixture of diastereomers 4 (0.785 g, 81%) as an oil. LC-MS[M+H]⁺=620.3 (Theoretical, C₃₇H₃₈N₃O₆ ⁺=620.2).

Synthesis of (2S,3R)-methyl3-amino-2-(4-((R)-5-morpholinohexa-1,3-diynyl) benzamido)butanoate and(2S,3R)-methyl3-amino-2-(4-((S)-5-morpholinohexa-1,3-diynyl)benzamido)butanoate (5)

The mixture of diastereomers 4 (0.785 g, 1.27 mmol) was dissolved in DMF(13 mL) and morpholine (2.2 mL, 25.4 mmol) was added in one portion atambient temperature and allowed to stir for 18 h. The resultant slurrywas vacuum filtered and the filtrate was concentrated in vacuo. Thecrude solid was purified by flash chromatography (0-10% MeOH/CH₂Cl₂) toprovide the desired mixture of diastereomers 5 (0.307 g, 61%) as a paleyellow oil. LC-MS [M+H]⁺=398.2 (Theoretical, C₂₂H₂₈N₃O₄ ⁺=398.2).

Synthesis ofN-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((R)-5-morpholinohexa-1,3-diynyl)benzamide(92-1) andN-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-44(S)-5-morpholinohexa-1,3-diynyl)benzamide(92-2)

The mixture of diastereomers 5 (0.307 g, 0.722 mmol) was dissolved inisopropyl alcohol (8 mL) and the hydroxylamine solution (1.02 mL, 15.4mmol) was added in one portion at ambient temperature. To this was thenadded solid potassium hydroxide (0.130 g, 2.31 mmol) in one portion andthe solution was allowed to stir for 18 h at ambient temperature. Thevolatiles were removed in vacuo and the resulting crude material waspurified by reverse phase HPLC (5-40% MeCN/H₂O, w/0.1% TFA) thenlyophilized to dryness to provide both of the desired products 92-1(0.0xx g, xx %) as a white solid; LC-MS [M+H]⁺=399.1 (Theoretical,C₂₁H₂₇N₄O₄ ⁺=399.2). ¹H NMR (DMSO) δ 11.31 (s, 1H), 9.42 (br s, 1H),8.60 (d, J=8.3, 1H), 8.36 (br s, 3H), 7.91 (d, J=8.2, 2H), 7.71 (d,J=8.0, 2H), 4.33 (q, J=6.6, 1H), 3.60-3.70 (m, 6H), 2.55-2.65 (m, 4H),1.37 (d, J=6.7, 3H), 1.15 (d, J=8.3, 3H) and 92-2 (0.0xx g, xx %) as awhite solid; LC-MS [M+H]⁺=399.0 (Theoretical, C₂₁H₂₇N₄O₄ ⁺=399.2). ¹HNMR (DMSO) δ 11.15 (s, 1H), 8.87 (d, J=8.6, 1H), 8.05 (br s, 3H), 7.96(d, J=8.3, 2H) 7.72 (d, J=8.0, 2H) 4.39-5.41 (m, 2H), 3.78 (br s, 4H),3.56-3.59 (m, 1H) 2.97-3.09 (m, 4H), 1.49 (d, J=5.9, 3H), 1.20 (d, 6.4,3H).

Example 93N-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((4-(methylsulfonamido)phenyl)buta-1,3-diynyl)benzamide(93-1)

Synthesis of methyl 4-((4-aminophenyl)buta-1,3-diynyl)benzoate (1)

The methyl 4-(2,2-dibromovinyl)benzoate (2.73 g, 8.54 mmol) and4-ethynylaniline (1.00 g, 8.54 mmol) were dissolved in DMF (40 mL) andthis solution was deoxygenated by bubbling N₂ through for 10 min. Tothis deoxygenated solution was then added Et₃N (3.6 mL, 25.6 mmol). N₂was then bubbled through for an additional 5 min followed by addition ofPd₂(dba)₃ (0.078 g, 0.085 mmol) and tri(4-methoxyphenyl)phosphine (0.121g, 0.342 mmol). The dark brown solution was then allowed to stir at 80°C. for 18 h then cooled to ambient temperature. The solution was dilutedwith EtOAc (100 mL), washed with deionized water (3×50 mL) and brine(1×50 mL), dried (MgSO₄), filtered and concentrated in vacuo. The crudeoil was purified by flash chromatography (0-50% EtOAc/CH₂Cl₂) to providethe desired product 1 as a yellow solid (0.671 g, 29%). LC-MS[M+H]⁺=276.0 (Theoretical, C₁₈H₁₄NO₂ ⁺=276.1).

Synthesis of methyl4-((4-(methylsulfonamido)phenyl)buta-1,3-diynyl)benzoate (2)

To a solution of compound 2 (0.370 g, 1.34 mmol) in CH₂Cl₂ (13 mL) wasadded methanesulfonyl chloride (0.11 mL, 1.47 mmol) slowly, followed bydropwise addition of Et₃N (0.21 mL, 1.47 mmol). The solution was allowedto stir at ambient temperature for 3 h. The solution was diluted withEtOAc (50 mL) and washed with aqueous HCl (1.0 N, 1×25 mL), aqueousNaHCO₃ (sat., 1×25 mL) and brine (1×25 mL). The organics were then dried(MgSO₄), filtered and concentrated in vacuo to provide the desiredproduct 2 (0.345 g, 73%) as an orange solid. ¹H NMR (DMSO) δ 7.99 (d,J=7.3, 2H), 7.72-7.80 (m, 4H), 7.62 (d, J=7.5, 2H), 3.87 (s, 3H), 2.40(s, 3H).

Synthesis of 4-((4-(methylsulfonamido)phenyl)buta-1,3-diynyl)benzoicacid (3)

To a solution of compound 2 (0.345 g, 0.976 mmol) in a mixture of THF (3mL), MeOH (3 mL), and water (3 mL) was added LiOH monohydrate (0.123 g,2.93 mmol) in one portion. The solution was allowed to stir at ambienttemperature for 90 min. The pH of the solution was adjusted with aqueousHCl (1.0 N) to pH˜1. The resulting solution was then extracted withEtOAc (3×20 mL), dried (MgSO₄), filtered and concentrated in vacuo thedesired product 3 (0.176 g, 53%) as a yellow solid. ¹H NMR (DMSO) δ10.24 (s, 1H), 7.96 (d, J=7.9, 2H), 7.71 (d, J=8.0, 2H), 7.60 (d, J=8.4,2H), 7.23 (d, J=8.5, 2H), 3.09 (s, 3H).

Synthesis of (2S,3R)-methyl3-(tert-butoxycarbonylamino)-2-(4-((4-(methylsulfonamido)phenyl)buta-1,3-diynyl)benzamido)butanoate (4)

Reagent MW Eq. g/mL mmol Compound 3 339.37 1.0 0.176 g 0.518(2S,3R)-methyl 3-(Boc)-2- 268.74 1.1 0.153 g 0.570 aminobutanoate EDC191.70 2.0 0.199 g 1.04 HOBT 135.12 2.0 0.141 g 1.04N,N-Diisopropylethylamine 129.24 4.0 0.34 mL 2.08 CH₂Cl₂ 5 mL

Compound 4 (0.285 g, 99%) as a yellow solid was prepared using GeneralEDC coupling. ¹H NMR (DMSO) δ 10.23 (s, 1H), 8.44 (d, J=8.2, 1H), 7.92(d, J=8.4, 2H) 7.74 (d, J=8.4, 2H), 7.60 (d, J=8.6, 2H), 7.23 (d, J=8.6,2H) 7.00 (d, J=9.7, 1H), 4.62 (dd, J=4.1, 8.5, 1H) 4.22-4.25 (m, 1H)3.61 (s, 3H), 3.09 (s, 3H), 1.19 (s, 9H), 1.11 (d, J=6.8, 3H).

Synthesis of (2S,3R)-methyl3-amino-2-(4-((4-(methylsulfonamido)phenyl)buta-1,3-diynyl)benzamido)butanoate(5)

To a solution of compound 4 (0.285 g, 0.515 mmol) in of CH₂Cl₂ (10 mL)was added TFA (2 mL) slowly. The solution was allowed to stir at ambienttemperature for 1 h then concentrated in vacuo. The thick oil was thenazeotroped with CH₂Cl₂ (3×) to provide the TFA salt of the desiredproduct 5 (0.284 g, 100%) as an off white solid. LC-MS [M+H]⁺=454.1(Theoretical, C₂₃H₂₄N₃O₅S⁺=454.1).

Synthesis ofN-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((4-(methylsulfonamido)phenyl)buta-1,3-diynyl)benzamide 93-1

The TFA salt of 5 (0.284 g, 0.515 mmol) was dissolved in isopropylalcohol (6 mL) and the hydroxylamine solution (0.68 mL, 10.3 mmol) wasadded in one portion at ambient temperature and the solution was allowedto stir for 18 h at ambient temperature. The volatiles were removed invacuo and the resulting crude material was purified by reverse phaseHPLC (5-40% MeCN/H₂O, w/0.1% TFA) then lyophilized to dryness to providethe desired product (93-1) (0.067 g, 29%) as a white solid; LC-MS[M+H]⁺=455.1 (Theoretical, C₂₂H₂₃N₄O₅S⁺=455.1).

Example 94 Synthesis of4-(cyclopropylbuta-1,3-diynyl)-N-(2-(hydroxyamino)-1-(1-hydroxycyclopropyl)-2-oxoethyl)benzamide94-1

Synthesis of 1-(2,2-dimethoxyethyl)cyclopropanol (1)

A solution of methyl 3,3-dimethoxypropanoate (28.2 g, 190 mmol) in athree-neck flask equipped with a overhead stirrer was cooled to 0° C.under N₂. The Ti(OiPr)₄ (11.3 mL, 38.1 mmol) was then slowly added andthe solution was allowed to stir for 10 min at 0° C. To this cooledsolution with vigorous stirring was then added the EtMgBr solution (158mL, 475 mmol) via syringe pump at ˜3 mL/min. After the addition wascomplete, the solution was allowed to slowly warm to ambient temperatureand stirred for an additional 18 h. The solution was then cooled to 0°C. and deionized water (45 mL) was slowly added with vigorous stirring.After stirring for 20 min at 0° C. the mixture vacuum filtered through apad of anhydrous MgSO₄. The clear, colorless solution was theconcentrated in vacuo and purified by flash chromatography (0-50%EtOAc/Hex) to provide the desired product (1) as a clear, colorless oil(8.60 g, 31%). ¹H NMR (DMSO) δ 5.04 (s, 1H), 4.62 (t, J=5.3, 1H), 3.22(s, 6H), 1.69 (d, J=5.3, 2H) 0.50 (dd, J=4.9, 6.9, 2H), 0.37 (dd, J=4.2,6.2, 2H).

Synthesis of 1-(2,2-dimethoxyethyl)cyclopropyl acetate (2)

To a solution of compound 1 (8.60 g, 58.8 mmol) in CH₂Cl₂ was added DMAP(0.718 g, 5.88 mmol) and pyridine (7.13 mL, 88.2 mmol) sequentially. Thesolution was then cooled to 0° C. and acetic anhydride was addeddropwise. After the addition was complete, the solution was stirred at0° C. for 10 min then allowed to warm to ambient temperature and stirredfor 18 h. The solution was then washed with aqueous NaHSO₄ (1.0 N, 2×100mL), aqueous NaHCO₃ (sat., 2×100 mL), dried (MgSO₄), filtered andconcentrated in vacuo. Isolated the desired product, (2) as a clear,colorless oil (11.1 g, 99%). ¹H NMR (DMSO) δ 4.50 (t, J=5.5, 1H), 3.21(2, 6H), 1.95 (d, J 5.3, 2H), 1.92 (s, 3H), 0.73-0.78 (m, 2H), 0.66-0.71(m, 2H).

Synthesis of 2-(1-acetoxycyclopropyl)acetic acid (3)

Compound 2 (11.1 g, 58.8 mmol) was dissolved in THF (50 mL) and water(100 mL) and cooled to 0° C. To this cooled solution was added Oxone® inportions. After complete addition the slurry was allowed to stir at 0°C. for 10 min the warmed to ambient temperature. After 8 h at ambienttemperature the slurry was diluted with deionized H2O (100 mL) andextracted with EtOAc (3×150 mL). The combined organics were dried(MgSO₄), filtered and concentrated in vacuo. The product (3) wasisolated as a thick, colorless oil (9.1 g, 98%). ¹H NMR (DMSO) δ 2.66(s, 2H), 1.92 (s, 3H), 0.77-0.85 (m, 4H).

Synthesis of methyl 2-(1-acetoxycyclopropyl)-2-bromoacetate (4)

Compound 3 (3.35 g, 21.2 mmol) was dissolved in anhydrous DCE (60 ml)under N₂ and cooled to 0° C. Thionyl chloride (3.5 g, 30 mmol) was thenadded slowly and the solution was refluxed for 30 min. At ambienttemperature, were added NBS (4.78 g, 26 mmol) and 4 drops ofconcentrated HBr and the solution was heated under reflux for 4 h. MeOH(50 ml) was added at room temperature and the solution was stirred at rtfor 3 h. The solvent were removed under reduced pressure, water (100 ml)was added and product was extracted with ethyl acetate, dried (Na2SO4),and concentrated to give 3.8 g of 4.

Synthesis of methyl 2-amino-2-(1-hydroxycyclopropyl)acetate (5)

The bromide product 4 was redessolved in dry DMF (20 ml), NaN₃ (2.05 g,30 mmol) was added and heated at 85° C. for 4 h. The reaction mixturewas diluted with ethyl acetate (100 ml) and water 5 ml was added. Theorganic layer was separated, dried and concentrated. The product (2.6 g)was the hydrogenated Pd—C (1.2 g) (40 psi) for 2 h. The catalyst wasfiltered off and ethanol solution were concentrated to give 5 (2.42 g)

Synthesis of methyl2-(4-(cyclopropylbuta-1,3-diynyl)benzamido)-2-(1-hydroxycyclopropyl)acetate(7)

Reagent MW Eq. g/ml mmol 4-(cyclopropylbuta-1,3- 201 1 210 mg 1diynyl)benzoic acid methyl 2-amino-2-(1-hydroxy- 145 2 300 mg 2.06cyclopropyl)acetate HATU 380.23 1.0 395 mg 1.05 DIEA 129.24 4 1.2 mLexcess CH3CN 10 ml

Compound (7) (172 mg) was prepared using the General Method for HATUcoupling and used on next step without purification. LC-Ms (M+1) 338.

Synthesis of4-(cyclopropylbuta-1,3-diynyl)-N-(2-(hydroxyamino)-1-(1-hydroxycyclopropyl)-2-oxoethyl)benzamide(94-1)

Aq. Hydroxylamine (2 ml, 50% aq.) was added to a stirred solution ofester 7, (0.17 g, 0.51 mmol) in isopropanol (15 ml), stirred for 18 h.Excess solvent was removed and the product was purified on a reversephase HPLC to give compound (94-1) (42 mg) LC-MS (M+1) 339: ChemicalFormula: C19H18N2O4, MW: 338.36.

Example 95

Example 95 was prepared by using the General Method for HATU coupling,General Method for Boc deprotection and General Method for hydroxamateformation

Synthesis of Intermediate Acid IC-3

(1s, 4s)-methyl 4-hydroxycyclohexanecarboxylate 2

Methanol (104 mL) was cooled in an ice-NaCl bath and SOCl₂ (15.2 mL) wasadded dropwise. To the resultant solution of HCl in methanol, compound 1(15 g, 0.1042 mol, 1.0 equiv) was added and the reaction mixture wasrefluxed for 1 h. After that time, Methanol was evaporated and another104 mL of a 2M solution of HCl in MeOH, prepared in the same way asbefore, was added and the reaction mixture was refluxed for anotherhour. Methanol was evaporated again to give compound 2 (16.3 g, 99%) asa light yellow liquid. ¹H NMR (400 MHz, DMSO-d₆): 1.47-1.83 (m, 9H),2.36-2.51 (m, 1H), 3.63 (s, 3H), 4.26 (s, 1H)

(1s,4s)-Methyl 4-(tert-butyldimethylsilyloxy) cyclohexanecarboxylate (3)

To a solution of compound 2 (16.3 g, 0.1032 mol, 1.0 equiv) andimidazole (14.03 g, 0.206 mol, 2.0 equiv) in dried DMF (100 mL) wasadded TBSCl (17.1 g, 0.114 mol, 1.1 equiv) in eight portions at 0° C.,then the reaction mixture was warmed to 30° C. and reacted at thistemperature for 2 h. DMF was evaporated, DCM (150 mL) was added, washedwith water (80 mL×2) and brine (100 mL), dried over anhydrous sodiumsulfate, evaporated the DCM and purified by silica-gel column (EA:PE,1:50) to give compound 3 (17 g, 61%) as a colorless liquid. ¹H NMR (400MHz, CDCl₃): δ 0.032 (s, 6H), 0.88 (s, 9H), 1.45-1.51 (m, 2H), 1.61-1.68(m, 4H), 1.90-1.98 (m, 2H), 2.29-2.34 (m, 1H), 3.67 (s, 3H), 3.89 (s,1H)

(1s, 4s)-4-(tert-butyldimethylsilyloxy) cyclohexyl) methanol (4)

To a stirred suspension of LiAlH₄ (1.9 g, 0.05 mol) in dried THF (50mL), under N₂, cooled to 0° C., a solution of compound 3 (17 g, 0.0625mol) in dried THF (20 mL) was added dropwise. The reaction mixture wasstirred for 1.5 h at 0° C., then quenched by the addition ofNa₂SO₄.10H₂O (5 g), filtered and evaporated the solvent, purified bysilica-gel column (EA: PE, 1:20) to give compound 4 (10.5 g, 69%) as athick oil. ¹H NMR (400 MHz, DMSO-d₆): δ 0.02 (s, 6H), 0.86 (s, 9H),1.14-1.41 (m, 7H), 1.51-1.55 (m, 2H), 3.17 (t, J=10.8 Hz, 2H), 3.94 (s,1H), 4.36 (t, J=10.8 Hz, 1H)

(1s, 4s)-4-(tert-butyldimethylsilyloxy) cyclohexanecarbaldehyde (5)

IBX (14.45 g, 0.052 mol) was dissolved in DMSO (50 mL) completely, thencompound 4 (10.5 g. 0.043 mol) was added in one portion and reacted at30° C. for 3 h. Water (60 mL) was added to quench the reaction,filtered, EA (100 mL) was added, washed with water (80 mL×2), brine (100mL), dried over anhydrous sodium sulfate, evaporated EA and the residuewas purified by silica-gel column (EA:PE, 1:50) to give compound 5 (6.29g, 50%) as a colorless liquid. ¹H NMR (400 MHz, DMSO-d₆): 0.021 (s, 6H),0.84 (s, 9H), 1.43-1.48 (m, 2H), 1.51-1.60 (M, 4H), 1.67-1.76 (m, 2H),2.27-2.33 (m, 1H), 3.86 (s, 1H), 9.56 (s, 1H)

Tert-butyl ((1s, 4s)-4-ethynylcyclohexyloxy) dimethylsilane (6)

To a solution of compound 5 (6.29 g, 0.026 mol) and K₂CO₃ (7.18 g, 0.052mol) in methanol (50 mL) was addeddimethyl-1-diazo-2-oxopropylphosphonate (5.99 g, 0.031 mol) and stirredovernight. The reaction mixture was diluted with Et₂O (50 mL), washedwith an aqueous solution (5%) of NaHCO₃ (30 mL) and dried over anhydroussodium sulfate, after filtration and evaporation of the solvent in cacuoto give compound 6 (3.09 g, 50%) as a light yellow liquid. ¹H NMR (400MHz, DMSO-d₆): 0.025 (s, 6H), 0.85 (s, 9H), 1.16-1.36 (m, 3H), 1.51 (d,J=4.8 Hz, 2H), 1.74-1.87 (m, 3H), 2.26-2.30 (m, 1H), 2.83-2.86 (m, 1H),3.6-3.74 (m, 1H)

Methyl 4-(((1s, 4s)-4-(tert-butyldimethylsilyloxy) cyclohexyl) buta-1,3-diynyl) benzoate (7)

Reagent MW Eq. mmol g, mL Compound 6 238.44 1.3 13 3.09 g Methyl4-(bromoethynyl)benzoate 239.07 1.0 10 2.58 g Pd(PPh₃)₂Cl₂ 701.9 0.050.5 0.38 g CuI 190.45 0.05 0.5 0.1 g Diisopropylamine 101.19 3 30 4.54mL THF 50 mL

Compound 7 (2.9 g, 58%) as a light yellow solid was prepared using thesame method for making IC-2. ¹H NMR (400 MHz, DMSO-d₆): 0.032 (s, 6H),0.85 (s, 9H), 1.23-1.95 (m, 8H), 2.57-2.73 (m, 1H), 3.59-3.77 (m, 1H),3.86 (s, 3H), 7.67 (d, J=8 Hz, 2H), 7.95 (d, J=8.4 Hz, 2H).

4-(((1s, 4s)-4-(tert-butyldimethylsilyloxy)cyclohexyl)buta-1,3-diynyl)benzoic acid (8)

Reagent MW Eq. mmol g, mL Compound 7 396.59 1.0 7.32 2.9 g NaOH (2Nsolution) 7 mL THF 7 mL Methanol 7 mL

Compound 8 (2.0 g, 71%) as a white solid made and separated using theGeneral Method for basic hydrolysis. ¹H NMR (400 MHz, DMSO-d₆): 0.036(s, 6H), 0.85 (s, 9H), 1.28-1.95 (m, 8H), 2.57-2.77 (m, 1H), 3.65-3.78(m, 1H), 7.64 (d, J=8 Hz, 2H), 7.93 (d, J=8.4 Hz, 2H), 13.23 (s, 1H)

Synthesis of Intermediate Acid IC-4

Diethyl tetrahydropyran-4,4-dicarboxylate (1)

Under ice bath, a three-necked flask was added 600 mL EtOH, then 23 g (lmmol) Na was added. After Na was almost dissolved, diethyl malonate 160ml (1 mol) and 1-chloro-2-(2-chloroethoxy)ethane (118 ml,1.2 mol) wereadded dropwise sequently to the solution at R.T. and stirred for anotherhalf hour. Then the mixture was heated to reflux for 1 h and cooled tor.t., then the other part of NaOEt (25 g Na dissolved in 600 mL EtOH)solution (1.2 mol) was poured into the mixture and refluxed overnight.The next day, the solvent was evaporated. The residue was dissolved inwater and extracted the product with EtOAc three times. The organiclayer was washed with brine, dried on Na₂SO₄ and evaporated to give thecrude product, which gave the pure product (120 g, Y=52%) bydistillation. ¹H NMR (400 MHz, CDCl₃): δ ppm 4.2 (q, 4H), 3.68 (m, 4H),2.12 (m, 4H), 1.26 (t, 6H).

Tetrahydropyran-4,4-dicarboxylic acid (2)

To a solution of compound 1 (40.0 g, 1.0 equiv) in CH₃OH/THF/H₂O=5/5/1(600 mL) was added NaOH (27.8 g, 4.0 equiv) and the mixture reacted at25° C. overnight. Then, the reaction solvents was removed andneutralized with 1 N HCl to PH=3-5. The product was extracted with EtOAcand concentrated under reduced pressure to give the desired product 2(27.3 g 91%). ¹H NMR (400 MHz, DMSO-d₆): δ ppm 12.84 (S, 2H), 3.53 (M,4H), 1.89 (m, 4H).

Tetrahydro-2H-pyran-4-carboxylic acid (3)

2 (27.3 g) was placed in a bottle and gradually heated to 170° C. over aperiod of 1 h until gas was ceased. The left solid was the desiredproduct 3 (20.4 g, Y=100%). ¹H NMR (400 MHz, DMSO-d₆): δ ppm 12.19 (s,1H), 3.80 (m, 2H), 3.28 (m, 2H), 2.47 (m, 1H), 1.69 (m, 2H), 1.53 (m,2H).

(Tetrahydro-2H-pyran-4-yl)methanol (4)

3 (6.5 g, 50 mmol. 1 eq.) was dissolved in dry THF (150 mL) and addedBF₃.Et₂O (7.0 mL, 50 mmol, 1.0 eq.) dropwise. The mixture was heated toreflux. While at this temperature 1M/L BH₃.THF (55 mL, 55 mmol, 1.1 eq.)was added dropwise. When the addition finished, the mixture was leftrefluxing overnight. The next day 3M/L NaOH solution □970 mL. 4.0 eq.)was added while the mixture was cooled to r.t. and reheated to refluxfor 1.5 h. Later, solvents were removed and the product was extractedwith EtOAc three times. The organic layer was washed with brine driedand evaporated to give the crude product which purified by columnchromatography (PE/EA=6:1-2:1) to obtain the pure desired product 1 4(4.8 g, Y=83%). ¹H NMR (400 MHz, DMSO-d₆): δ ppm 4.31 (m, 1H), 3.71 (m,2H), 3.09 (m, 4H), 1.40 (m, 3H), 0.98 (m, 2H).

Tetrahydro-2H-pyran-4-carbaldehyde (5)

IBX (5.8 g, 1.2 eq.) were dissolved in 30 mL DMSO at 40° C. and 4 (2.0g, 1.0 eq.) was added to the mixture and left stirring overnight. Thenext day, filtrated the solid, the filtrate was diluted with 80 mLwater, and extracted the product with ether four times. The ether layerwas washed with brine dried and evaporated the solvents under ice bathto give the crude product 1.3 g (the product was broken up under theair). ¹H NMR (400 MHz, DMSO-d₆): δ ppm 9.62 (s, 1H), 3.81 (m, 2H), 3.34(m, 2H), 2.56 (m, 1H), 1.75 (m, 2H), 1.50 (m, 2H).

4-(2,2-Dibromovinyl)-tetrahydro-2H-pyran (6)

To a 0° C. solution of compound 18-5 (1.3 g, 11.4 mmol, 1.0 eq.) andcarbon tetrabromide (4.2 g, 12.5 mol, 1.1 eq.) in dry CH₂Cl₂ (30 ml) wasadded triphenylphosphine (6.27 g, 24.0 mmol, 2.1 equiv.) in 4 portionsat 3 min intervals. The reaction was then stirred for 1 h at 25° C.Hexane was added to the reaction mixture with good stirring, and theresulting slurry was filtered through silica gel and evaporated to givethe crude product, which purified by column chromatography (PE/EA=0-10%)to obtain the pure product 6 (2.0 g, 65%). ¹H NMR (400 MHz, DMSO-d₆): δppm 6.20 (d, J=8.8 Hz), 3.87 (m, 2H), 3.37 (m, 2H), 2.47 (m, 1H), 1.57(m, 2H), 1.42 (m, 2H).

Methyl 4-((tetrahydro-2H-pyran-4-yl)buta-1,3-diynyl)benzoate (7)

A solution of 6 (2.0 g 7.4 mmol, 1 equiv), methyl 4-ethynylbenzoate(1.80 g 11.1 mmol, 1.5 equiv), Pd₂dba₃ (68 mg, 0.08 mmol, 0.01 equiv),TMPP (106 mg, 0.32 mmol, 0.04 equiv) and TEA (3 mL, 22.2 mmol, 3 equiv)in anhydrous DMF (30 ml) was flushed with N₂ and heated at 80° C. for 10hours. Then, most DMF was evaporated under reduced pressure, the residuewas dissolved in EtOAc (500 ml) and washed with semi-saturated NaClsolution (300 ml×4), dried on Na₂SO₄ and evaporated to give a crudeproduct, which was purified by chromatography (EA/PE=1/20-1/5) to givethe desired product 7 (360 mg, 30%). MS: [M+1]⁺269

4-((Tetrahydro-2H-pyran-4-yl)buta-1,3-diynyl)benzoic acid (IC-4)

Reagent MW Eq. mmol g, mL Compound 7 268 1.0 1.3 360 mg NaOH 40 4.0 5.4300 m CH₃OH/THF/H₂O = 5/5/1 50 mL

Compound IC-4 (310 mg, 98%) was made and separated using the GeneralMethod for basic hydrolysis. MS: [M+1]⁺255. ¹H NMR (400 MHz, DMSO-d₆): δppm 13.21 (s, 1H), 7.94 (d, 0.1=8.4 Hz, 2H), 7.66 (d, J=8.4 Hz, 2H),3.77 (m, 2H), 3.44 (m, 2H0, 2.93 (m, 1H), 1.83 (m, 2H), 1.62 (m, 2H).

Synthesis of Intermediate Acid IC-5

Methyl 4-(2,2-difluorocyclopropyl buta-1,3-diynyl)benzoate (5)

IBX (2.8 g, 10 mmol) was added to DMSO (20 mL) in one portion. After IBXwas totally dissolved, compound 1 was added in one portion and themixture was stirred for 3 h at 25° C. Then, 80 mL water was added andthe white precipitate was filtered off without suction. The filtrate wasextracted with toluene (3×15 mL). Then the organic layer was washed withwater (5×10 mL), dried overnight with anhydrous MgSO₄. MgSO₄ wasfiltered off and the solid was washed twice with toluene. A 3-neck flaskcharged with K₂CO₃ (2.61 g, 18 mmol) was flushed with nitrogen 3 times.Then, MeOH (30 mL), the toluene solution from the former step anddimethyl-1-diazo-2-oxopropyl phosphonate (1.728 g, 9 mmol) were addedsequentially. The reaction mixture was stirred at r.t. overnight andwashed with distilled water (3×10 mL). Then, the organic layer was driedover anhydrous MgSO₄ for 4 hours. Filtration without suction gave thetoluene solution of compound 3 for the next step. Another 3-neck flaskwith methyl 4-ethynyl benzoate (480 mg, 3 mmol), Pd(PPh₃)₂Cl₂ (105 mg,0.15 mmol), CuI (57 mg, 0.3 mmol) was charged with N₂. The dried toluenesolution with compound 3 in it was put into the flask by a syringe.Diisopropylamine (909 mg, 9 mmol) was added dropwise. The reactionmixture was stirred overnight and filtered. The solid was washed withEtOAc (3×20 mL). The filtrate was concentrated in vacuo and purified byflash chromatography (PE:EA=50:1) to give compound 5 (262 mg, yield 11%from 3 steps) as a yellow solid. ¹H NMR (500 MHz, CDCl₃): δ ppm, 7.98(d, J=8.0 Hz, 2H), 7.54 (d, J=8.0 Hz, 2H), 3.92 (s, 3H), 2.25-2.32 (m,1H), 1.79-1.83 (m, 1H), 1.63-1.68 (m, 1H).

4-(2,2-Difluorocyclopropyl buta-1,3-diynyl)benzoic acid (IC-5)

To a solution of compound 5 (260 mg, 1 mmol) in THF (5 mL), MeOH (5 mL)and a solution of NaOH (160 mg, 4 mmol) in water (2 mL) was added at 0°C. Then, the mixture was stirred at r.t. for 2 h as monitored by TLC.The solvent was evaporated and 20 mL water was added. Then, the mixturePH value was adjusted to 2-3 and extracted with EtOAc (3×15 mL). Theorganic layer was dried over MgSO₄, filtered and concentrated to givecompound IC-5 (200 mg, 81%) as a light pink solid. ¹H NMR (500 MHz,DMSO-d₆): δ ppm, 7.94 (d, J=6.5 Hz, 2H), 7.69 (d, J=6.5 Hz, 2H),2.90-2.94 (m, 1H), 2.15-2.19 (m, 1H), 1.90-1.97 (m, 1H).

Synthesis of Intermediate Acid IC-6

Step 1: Trimethyl(4-methylpent-3-en-1-ynyl)silane 2

Under nitrogen, ethynyltrimethylsilane 1 (3.5 g, 0.036 mol, 1.2 equiv)was added to a solution of compound SM1 (4 g, 0.03 mol, 1.0 equiv),PdCl₂(PPh₃)₂ (1.05 g, 1.5 mmol, 0.05 equiv) and CuI (0.6 g, 0.003 mmol,0.1 equiv) in TEA (50 mL) at 90° C. The mixture was allowed to reactovernight. Then, the precipitate was isolated. The filter cake waswashed with Et₂O (100 mL). The filtrate was concentrated under reducedpressure. The residue was dissolved in Et₂O (100 mL) and washed 1 M HClaq. (200 mL), water (200 mL×2) and brine (200 mL). The organic layer wasseparated, dried (Na₂SO₄) and the filtered. The filtrate wasconcentrated under reduced pressure to give the crude product of titlecompound 2 as liquid (3 g, 65% yield). The purity was about 70% fromLCMS.

Step 2: Methyl 4-(6-methylhepta-5-en-1,3-diynyl)benzoate 3

To a solution of compound 2 (2 g, 0.02 mol, 1.0 equiv) in DMF (50 mL)was treated with K₂CO₃ (16 g, 0.12 mol, 4 equiv) at r.t. Then, themixture was stirred at room temperature for 1 h. Then, the reactantmethyl 4-(bromoethynyl)benzoate SM2 (5 g, 0.02 mol, 1.0 equiv),PdCl₂(PPh₃)₂ (0.7 g, 0.001 mol, 0.05 equiv) and CuI (0.38 g, 0.002 mmol,0.1 equiv) and diisopropylamine HN(i-Pr)₂ (4 g, 0.04 mol, 2.0 equiv) wasadded in to the reaction mixture quickly under N₂ protection. Themixture was stirred for overnight. Then, the reaction mixture was addedEtOAc (200 mL) and washed with dilute HCl aqueous (150×2), H₂O (200×2).The organic layer was washed, dried (Na₂SO₄), filtered and concentratedunder reduced pressure. The residue was purified by chromatography onsilica gel to give the desired product 3 (0.4 g, 10%). LCMS (m/z):[M+H]⁺=239.0. ¹H NMR (300 MHz, CDCl₃): δ 7.97-7.99 (m, 2H), 7.57-7.49(m, 2H), 5.40 (s, 1H), 3.92 (s, 3H), 1.98 (s, 3H), 1.87 (s, 3H).

Step 3: 4-(6-Methylhepta-5-en-1,3-diynyl)benzoic acid 4

To a solution of compound 3 (0.25 g, 1.7 mmol, 1.0 equiv) in methanol (5mL) was treated with KOH/H₂O (0.3 g/3 mL) at r.t. The reaction mixturewas neutralized with acetic acid and EtOAc (100 mL). The organic layerwas washed with 5% aq. Na₂CO₃ (50 mL×2), dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The residue was dried in vacuoovernight to provide the title compound IC-6 (0.2 g, 87%). LCMS (m/z):[M+H]⁺=225.0. ¹H NMR (300 MHz, DMSO): δ 7.93-7.95 (d, 2H), 7.66-7.68 (d,2H), 5.58 (m, 1H), 1.93 (s, 3H), 1.87 (s, 3H).

Synthesis of Intermediate Acid IC-7

Methyl 4-(5-(methylsulfonyloxy)penta-1,3-diynyl)benzoate (2)

To a stirred solution of compound 1 (0.7 g, 3.27 mmol), TEA (0.55 mL,3.92 mmol) in dichloromethane (30 mL) and THF (10 mL) was addedmethanesulfonyl chloride (0.56 g, 0.38 mL, 4.9 mmol) dropwise under icecooling. After addition cooling-bath was removed and stirred continuedfor 2 hours. The mixture was diluted with dichloromethane (100 mL) andwashed successively with water (30 mL), saturated NH₄Cl (20 mL), brine(50 mL), dried (Na₂SO₄) and filtered. The residue was evaporated to givethe crude product as a dark brown solid. MS (m/z): [M+H]⁺=293

Methyl 4-(5-(dimethylamino)penta-1,3-diynyl)benzoate (3)

To a solution of compound 2 (0.533 g, 3.27 mmol) in DMF (20 mL), wasadded dimethylamine hydrochloride (3.2 g, 39 mmol), followed by DIPEA(5.9 g, 7.9 mL, 45.8 mmol). After being stirred at 65° C. for 2 hours,cooled down to room temperature, diluted with saturated NaHCO₃ aqueousand extracted with dichloromethane (3×100 mL). The combined organiclayers was washed (water, brine), dried (Na₂SO₄), filtered andevaporated to dry. The crude product was purified by silica gel columnchromatography (PE/EA=20:1) to give the desired product (0.2 g, 25%). MS(m/z): [M+H]⁺=242. ¹H-NMR (400 MHz, DMSO-d₆): 2.22 (s, 6H), 3.50 (s,2H), 3.59 (s, 3H), 7.72 (d, J=8.4 Hz, 2H), 7.96 (d, J=8.8 Hz, 2H).

4-(5-(Dimethylamino)penta-1,3-diynyl)benzoic acid (IC-7)

To a solution of compound 3 (0.2 g, 0.83 mmol) in a mixture solution ofTHF (20 mL) and CH₃OH (10 mL) was added 2N NaOH (5 mL) dropwise atambient temperature. The mixture was allowed to react at roomtemperature for 3 hours, distilled the solvent, extracted with ether.The aqueous layer was adjusted the PH at 7 with 2 N HCl aqueous andfiltered to give compound 4 (129 mg, 68%) as a yellow solid. MS (m/z):[M+H]⁺=228. ¹H-NMR (400 MHz, DMSO-d₆): 2.23 (s, 6H), 3.51 (s, 2H), 3.59(s, 3H), 7.69 (d, J=8 Hz, 2H), 7.94 (d, J=7.6 Hz, 2H).

Synthesis of Intermediate Acid IC-8

5-(5-Bromothiophen-2-yl)oxazole (2)

To a degassed solution of tosylmethyl isocyanide (0.56 g, 2.88 mmol),K₂CO₃ (0.4 g, 2.88 mmol) in dried CH₃OH (15 mL) was added compound 1(0.5 g, 2.9 mL, 2.6 mmol). The reaction mixture refluxed under nitrogenfor 2 hours. Volatiles were removed in vacuo and the residue waspurified by silica gel column (PE:EA=50:1) to give compound 2 (0.45 g,75%) as a light yellow solid. ¹H-NMR (400 MHz, CDCl₃): 7.04 (d, J=4 Hz,1H), 7.07 (d, J=4 Hz, 1H), 7.19 (s, 1H), 7.86 (s, 1H).

5-(5-((Trimethylsilyl)ethynyl)thiophen-2-yl)oxazole (3)

Under nitrogen ethynyltrimethylsilane (85 mg, 0.13 mL, 0.87 mmol) wasadded to a solution of compound 2 (0.1 g, 0.43 mmol), Pd(PPh₃)₂Cl₂ (10mg, 0.014 mmol), CuI (8 mg, 0.04 mmol) in TEA (5 mL) at 60° C. Thesolution was heated to 90° C. for 2 hours. The reaction was treated withEA (100 mL), washed with water (100 mL), brine (100 mL), dried (Na₂SO₄),evaporated in vacuo. The residue product was purified by silica gelcolumn (PE: EA=50:1) to give desired product (73 mg, 68%) as a lightyellow solid.

5-(5-Ethynylthiophen-2-yl)oxazole (4)

To a solution of compound 3 (4.3 g, 17.4 mmol) in a mixture solution ofmethanol (40 mL) and THF (50 mL) was treated with KOH/CH₃OH (0.49 g/10mL) dropwise below 10° C. The reaction mixture was allowed to react atambient temperature for 0.5 h. The reaction solution was neutralizedwith acetic acid to PH=7 and concentrated under reduced pressure. Theresidue was purified by silica gel column (PE: EA=50:1) to give desiredproduct (2.0 g, 67%) as a yellow solid. ¹H-NMR (400 MHz, CDCl₃): 3.44(s, 1H), 7.23 (d, J=2, 1H), 7.26 (d, J=1.6, 1H), 7.18 (s, 1H), 7.87 (s,1H),

Methyl 4-((5-(oxazol-5-yl)thiophen-2-yl)buta-1,3-diynyl)benzoate (5)

To a degassed solution of compound 4 (1.9 g, 10.8 mmol), methyl4-(bromoethynyl)benzoate (2.7 g, 11.4 mmol), Pd(PPh₃)₂Cl₂ (0.38 g, 0.54mmol), CuI (0.10 g, 0.54 mmol) in THF (90 mL) was added i-Pr₂NH (3.29 g,4.6 mL, 32.5 mmol) at room temperature. The mixture was allowed to reactat this temperature for 3 h. The mixture was treated with EA (300 mL),washed with water (200 mL), saturated NH₄Cl (100 mL), brine (300 mL),dried (Na₂SO₄), concentrated in vacuo. The residue was purified bysilica gel column (PE/EA=1:1) to give desired product (1.45 g, 40%) as ayellow solid.

4((5-(Oxazol-5-yl)thiophen-2-yl)buta-1,3-diynyl)benzoic acid (IC-8)

To a solution of compound 5 (1.43 g, 4.3 mmol) in a mixture solution ofTHF (300 mL) and CH₃OH (50 mL) was added 2 N NaOH (10 mL) at ambienttemperature. The reaction solution was allowed to react for 3 hours.Then evaporated the organic solvent, adjusted the PH at 6 with 2 N HCl.Filtration gave crude product, which was triturated in THF. Theprecipitate was collected by filtration to give desired product (0.262g, 20%) as a yellow solid. ¹H-NMR (400 MHz, DMSO-d₆): 7.52 (d, J=4, 1H),7.6 6 (d, J=3.6, 1H), 7.72-7.74 (m, 3H), 7.98 (d, J=8, 2H), 8.51 (s,1H).

Synthesis of Intermediate Acid IC-9

Methyl 4-(bromobuta-1,3-diynyl)benzoate (1)

0 (368 mg, 2 mmol, 1.0 eq.) and AgNO₃ (40 mg, 0.1 eq.) were dissolved in50 mL acetone and stirring for 2 h at r. t. Then, NBS (4 mmol, 2.0 eq.)was added to the mixture and stirred for 2 h. TLC monitored thereaction. When the reaction was complete, filtered the solid and thefiltrate was evaporated and dissolved in EtOAc. The organic layer waswashed with sat. NaHCO₃, H₂O and brine, dried, filtered and evaporatedto give the desired product 1 without purification (526 mg, Y=100%). ¹HNMR (400 MHz, CDCl3): δ ppm 8.00 (d, J=8.4 Hz, 2H), 7.56 (d, J=8.4 Hz,2H), 3.92 (s, 3H).

Methyl 4-(cyclopropylhexa-1,3,5-triynyl)benzoate (2)

Under nitrogen, diisopropylethylamine (DIPEA) (1 mL, 3.0 equiv) andethynylcyclopropane (4 mmol, 2.0 equiv.) were added to a solution ofcompound 1 (526 mg, 2 mmol 1.0 equiv), PdCl₂(PPh₃)₂ (0.181 g, 0.05 mmol,0.025 equiv) and CuI (0.191 g, 0.1 mmol, 0.1 equiv) in dry THF (50 mL)in ice bath. The mixture was allowed to react at r.t. overnight. Then,the solvents were evaporated and the residue was purified by columnchromatography to get the pure desired product 2 (380 mg, 76.6%). MS:[M+1]⁺=249

4-(Cyclopropylhexa-1,3,5-triynyl)benzoic acid (IC-9)

To a solution of compound 2 (380 mg, 1.0 equiv) in CH₃OH/THF/H₂O=5/5/1(50 mL) was added NaOH (300 mg, 4.0 equiv). Then, the mixture wasstirred at 25° C. overnight. Then, the solvent was removed andneutralized with 1 N HCl to PH=3-5. The product was extracted with EtOAcand concentrated under reduced pressure to give the desired product IC-9(310 mg, 86.6%). LC-MS: [M−H]⁻=233. ¹H NMR (400 MHz, DMSO-d₆): δ ppm13.32 (s, 1H), 7.95 (d, J=8.4 Hz, 2H), 7.73 (d, J=8.4 Hz, 2H), 1.61 (m,1H), 0.95 (m, 2H), 0.85 (m, 2H).

Synthesis of Intermediate Acid IC-10

The target product was prepared by following the General procedure forMethod 1-C (Cadiot-Chodkiewicz Coupling), Method 6 (DAST Fluorination)and Method 2-A (Basic hydrolysis)The following compounds were synthesized as described above.

MH⁺ Ret. HPLC- Compound (m/z) Time MS # Structure theo (min) Method95-1 

397 95-2 

397 95-3 

383 95-4 

396 95-5 

382 95-6 

398 95-7 

356 95-8 

368 95-9 

410 95-10

369 95-11

343 95-12

357 95-13

367 95-14

375 95-15

353 95-16

365 95-17

351 95-18

448 95-19

460 95-20

438 95-21

396 95-22

332 95-23

346 95-24

364 95-25

346

Example 96 Synthesis of(E)-4-[4-(4-morpholin-4-ylmethylphenyl)-but-3-en-1-ynyl]benzoic acid (1)

4-Morpholin-4-ylmethylbenzaldehyde (2)

To a solution containing morpholine (5.25 mL, 60 mmol) indichloromethane (50 mL) was added HCl (5 mL, 20 mmol, 4 M in dioxane)dropwise following by 4-diethoxymethylbenzaldehyde (1, 2.08 g, 10 mmol),NaBH₃CN (0.44 g, 7 mmol) and Aliquat 336 (2.93 g, 7 mmol). Then 1 g of 4A molecular sieves was added. The mixture was stirred at roomtemperature overnight, filtered, and the solvent removed in vacuo. Then200 mL water was added to the residue and the pH was adjusted to 4 by0.5 M citric acid. The mixture was stirred for 1 hour and then extractedwith ether (2×100 mL). The pH of the aqueous layer was adjusted to 9 by1.0 M sodium hydroxide. It was extracted with ether (3×100 mL). Thecombined organic layers were washed with water (150 mL), brine (50 mL)dried (MgSO₄), filtered and concentrated under reduced pressure to givethe title compound 2 (1.0 g, 48%). MS (m/z): [M+H]⁺=206. ¹H NMR (300MHz, CDCl₃): in ppm, 2.48 (br s, 4H), 3.59 (s, 2H), 3.70-3.78 (m, 4H),7.54 (d, J=8.0 Hz, 2H), 7.85 (d, J=8.3 Hz, 2H), 10.0 (s, 1H).

(E)-4-[4-(2-Iodovinyl)benzyl]morpholine (3)

Chromium chloride anhydrous (19.1 g, 155 mmol, 8 eq) was added to THF(400 mL) under nitrogen. A solution of triiodomethane (30.7 g, 78 mmol,4 eq) in THF (500 mL) was added dropwise at 0° C. under nitrogen. Then asolution of 2 (4.0 g, 19.5 mmol, 1 eq) in THF (100 mL) was addeddropwise. The mixture was stirred at 0° C. for 2 h and then at roomtemperature for 2 h. The mixture was poured into iced water andextracted with EtOAc (2×800 mL). The combined organic phase was washedwith 20% aq. Na₂S₂O₃ (300 mL), brine (300 mL), dried (MgSO₄), filteredand concentrated under reduced pressure. The residue was purified bychromatography on silica gel (0 to 5% EtOAc/Hexanes) to give the titlecompound 3 (3.55 g, 55%). MS (m/z): [M+H]+=330. ¹H NMR (300 MHz, CDCl₃):in ppm, 2.44 (t, J=4.6 Hz, 4H), 3.47 (s, 2H), 3.70 (t, J=4.6 Hz, 4H),6.80 (d, J=15 Hz, 1H), 7.22-7.27 (m, 4H), 7.41 (d, J=15 Hz, 1H).

(E)-4-[4-(4-morpholin-4-yl-methyl-phenyl)-but-3-en-1-ynyl]-benzoic acidmethyl ester (5)

To a mixture of 3 (290 mg, 0.88 mmol, 1.0 eq), 4 (141 mg, 0.88 mmol, 1.0eq.), diisopropylamine (0.125 mL, 0.88 mmol) and PdCl₂(PPh₃)₂ (32 mg,0.044 mmol, 0.05 eq) in Et₃N (15 mL) was added CuI (18 mg, 0.09 mmol,0.1 eq) under nitrogen at room temperature. The mixture was stirred atroom temperature for 12 h, diluted with EtOAc (40 mL), filtered andconcentrated. The residue was dissolved into EtOAc (50 mL) and washedwith water (50 mL) and brine (50 mL), and dried (MgSO₄). Afterfiltration and concentration, the residue was purified by chromatographyon silica gel (0 to 30% EtOAc/Hexanes) to give the title compound 5 (209mg, 66%). MS (m/z): [M+H]+=362. ¹H NMR (300 MHz, CDCl₃): in ppm, 2.44(t, J=4.6 Hz, 4H), 3.48 (s, 2H), 3.70 (t, J=4.6 Hz, 4H), 3.90 (s, 3H),6.36 (d, J=16.2 Hz, 1H), 7.06 (d, J=16.2 Hz, 1H), 7.30 (d, J=8.4 Hz,2H), 7.37 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.9 Hz, 2H), 7.99 (d, J=8.9 Hz,2H).

(E)-4-[4-(4-Morpholin-4-ylmethylphenyl)-but-3-en-1-ynyl]benzoic acid (1)

To a solution of 5 (4.3 g, 11.9 mmol, 1.0 eq) in THF/MeOH/H₂O (80 mL,1/1/1) was added LiOH.H₂O (1.0 g, 23.8 mmol, 2.0 eq). The mixture wasstirred at room temperature for 24 h. The pH was adjusted to 7 by 1 MHCl. The mixture was stirred for 1 h. The solid was filtered and washedwith water (40 mL) and ether (40 mL) and dried in vacuo. To a suspensionof above solid in ether (40 mL) was added HCl (24 mL, 48 mmol, 2 M inether). The mixture was stirred at room temperature for 1 h. The solidwas filtered and washed with ether (40 mL), and dried in vacuo at 45° C.overnight to give the target product 1-HCl (3.0 g, 71%). MS (m/z):[M+H]⁺=348. ¹H NMR (300 MHz, CDCl₃): in ppm, 2.95-3.20 (m, 4H),3.75-4.00 (m, 4H), 4.29 (s, 2H), 6.78 (d, J=16.2 Hz, 1H), 7.22 (d,J=16.2 Hz, 1H), 7.61 (d, J=8.4 Hz, 2H), 7.67 (br, s, 4H), 7.96 (d, J=8.4Hz, 2H), 11.7 (br s, 1H).

N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(96-1)

Synthesis of(2S,3R)-3-hydroxy-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-butyricacid methyl ester (2)

To the mixture of compound 1 (70 mg, 0.20 mmol) and H—(S)-Thr-OMehydrochloride (41 mg, 0.24 mmol) in DMF (1.5 mL) was added BOP (115 mg,0.26 mmol) followed by DIEA (104 μl, 0.6 mmol). Reaction mixture wasstirred at ambient temperature for 20 min, diluted with EtOAc (50 mL),extracted with water (30 mL×2) and brine (30 mL). Organic layer wasdried over anh. Na₂SO₄, evaporated in vacuo and dried in vacuo overnightto provide target product 2 (92 mg, 100%) as white solid. LC-MS [M+H]463.3 (C27H30N2O5+H, requires 463.55). Compound was used in nextsynthetic step without additional purification.

Synthesis ofN-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(96-1)

To a stirred suspension of compound 2 (92 mg, 0.2 mmol) andhydroxylamine hydrochloride (84 mg, 1.2 mmol) in MeOH (anh, 2 mL) andTHF (anh, 2 mL) was added NaOtBu (154 mg, 1.6 mmom) powder in oneportion at −5° C. under nitrogen. Reaction mixture was stirred at −5° C.for additional 5 min. Temperature of reaction mixture was raised toambient. Completion of the reaction was monitored by LC-MS. Aftercompletion (in 20 min) reaction mixture was acidified with 1 N HCl inMeOH to pH˜6 at low temperature and evaporated in vacuo. Residue wasdissolved in DMSO (600 μl) and subjected to HPLC purification.[Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flow rate=20 mL/min;mobile phase A: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1%TFA; gradient elution from 8% B to 28% B in 60 min., detection 254 nm].Fractions containing the desired product were combined and lyophilizedto provide trifluoroacetate salt of target product (15-1) (20.5 mg) aswhite solid. LC-MS [M+H] 464.3 (C26H29N3O5+H, requires 464.54).

Retention Scale Yield Yield Purity [M + time Compound (mmol)* (mg) (%)*(%) H] (min)** 96-1 0.2 20.5 17.8 97.7 464.3 4.06 *Based on the amountof carboxylic acid used in the coupling reaction **HPLC-MS Method BEach of the following compounds was synthesized as described above.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 96-2

464.4 4.08 B 96-3

462.4 5.01 B 96-4

463.3 3.57 B

Example 97N-((1S,2R)-1-Hydroxycarbamoyl-2-methoxy-propyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(97-1)

Synthesis of(2S,3R)-3-methoxy-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-butyricacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 1 (Ach-Tr-Db- 347.41 1.0 80 mg 0.23(Morpholine)-OH H-(S)-Thr(Me)-OTMS 133.15 1.2 37 mg 0.28 BSA 203.43 2.9160 μl 0.67 BOP 442.28 1.26 127 mg 0.29 DIEA 129.24 4.0 160 μl 0.92 DMF2 mL

Compound 3 (100 mg, 95%) as light yellow oil was prepared using the BOPcoupling described in Example 96 for compound 2. LC-MS [M+H] 463.2(C27H30N2O5+H, requires 463.5).

Synthesis of NN-[(1S,2R)-2-methoxy-1-(tetrahydro-pyran-2-yloxycarbamoyl)-propyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(4)

A solution of compound 3 (100 mg, 0.22 mmol), DIC (51 μl, 0.33 mmol) andHOAT (45 mg) in CHCl₃ (1.67 mL) and DMF (0.33 mL) was stirred at 0° C.for 10 min followed by addition of THP—O—NH₂ (52 mg, 0.44 mmol).Reaction mixture was stirred at 0° C. for additional 30 min. Thenreaction mixture was allowed to warm to ambient temperature, dilutedwith water (50 mL), extracted with CHCl₃ (50 mL×2). Combined organicphase was washed with brine (30 mL), dried over anh. Na₂SO₄ andconcentrated in vacuo. Flash chromatography on silica gel of the residuegave target product 4 (63 mg, 51%) as white solid. LC-MS [M+H] 562.4(C32H39N3O6+H, requires 562.7).

Synthesis ofN-((1S,2R)-1-hydroxycarbamoyl-2-methoxy-propyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(97-1)

A solution of compound 4 (63 mg, 0.11 mmol) in 0.5 mL of dioxane wascooled to 0° C. followed by 4 N HCl/dioxane (0.5 mL, 2 mmol). Reactionmixture was allowed to warm to ambient temperature and stirred for 30min. Solvent was evaporated in vacuo. Residue was dissolved in DMSO (400μl) and subjected to HPLC purification. [Phenomenex Gemini C-18 column,110 Å (30×100 mm); flow rate=20 mL/min; mobile phase A: 100% water, 0.1%TFA; mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to28% B in 60 min., detection 254 nm]. Fractions containing the desiredproduct were combined and lyophilized to provide trifluoroacetate saltof target product 97-1 (12.8 mg) as white solid. LC-MS [M+H] 478.2(C27H31N3O5+H, requires 478.7).

Retention Compound Scale Yield Yield Purity [M + time # (mmol)* (mg)(%)* (%) H] (min)** 97-1 0.23 12.8 9.4 95 478.2 4.8 *Based on the amountof carboxylic acid used in the coupling reaction **HPLC-MS Method BThe following compound was synthesized as described above.

HPLC- Compound MH⁺ Ret. Time MS # Structure (m/z) (min) Method 97-2

478.2 4.43 B

Example 98 Synthesis ofN—((S)-2-amino-1-hydroxycarbamoyl-ethyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(98-1)N—[(S)-2-(2-dimethylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(98-2)N—((S)-2-acetylamino-1-hydroxycarbamoyl-ethyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(98-3)

Synthesis of (S)-3-tert-butoxycarbonylamino-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (2)

Reagent MW Eq. mg/mL mmol Compound 1 (Ach-Tr-Db- 347.41 1.0 280 mg 0.80(Morpholine)-OH N-Boc-(S)-DAP-OMe × HCl 254.71 1.2 245 mg 0.96 BOP442.28 1.25 442.3 mg 1.0 DIEA 129.24 3.0 418 μl 2.4 DMF 5 mL

The product 2 (338 mg, 77%) as yellow solid was prepared using the BOPcoupling described in Example 96 for compound 2. LC-MS [M+H] 548.4(C31H37N3O6+H, requires 548.6). Compound was used in next synthetic stepwithout additional purification.

Synthesis of(S)-3-amino-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 2 547.64 1.0 338 mg 0.62 4NHCl/dioxane 13 2 mL 8 Dioxane/MeOH (1:1) 2 mL

The product 3 (321 mg, 100%) as white solid was made using the GeneralMethod for Boc deprotection. LC-MS [M+H] 448.4 (C26H29N3O4+H, requires448.5).

Synthesis ofN—((S)-2-amino-1-hydroxycarbamoyl-ethyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(98-1)

Reagent MW Eq. mg/mL mmol Compound 3 × 2 HCl 518.5 1.0 104 mg 0.2 NH₂OH× HCl 69.49 6.0  84 mg 1.2 NaOtBu 96.11 8.0 154 mg 1.6 MeOH 2 mL THF 2mL

The product 4 (20.5 mg) as white solid was prepared using theHydoxyamide formation described in Example 96 for compound 96-1. LC-MS[M+H] 449.2 (C25H28N4O4+H, requires 449.5).

Retention Scale Yield Yield Purity [M + time Compound (mmol)* (mg) (%)*(%) H] (min)* 98-1 0.2 20.5 18.3 97.5 449.2 3.48 *Based on the amount ofcompound 3. **HPLC-MS Method B

Synthesis of (S)-3-(2-dimethylamino-acetylamino)-2- {4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (5)

To the mixture of compound 3 di-hydrochloride (104 mg, 0.20 mmol) anddimethylaminoacetyl chloride hydrochloride (41 mg, 0.26 mmol) in DMF(0.5 mL) and CHCl₃ (1.5 mL) was added DIEA (210 μl, 1.2 mmol). Reactionmixture was stirred at ambient temperature overnight. Solvent wasconcentrated in vacuo. Residue was dissolved in EtOAc (50 mL), washedwith 5% solution of NaHCO₃ (50 mL) and brine (30 mL) Organic phase wasdried over anh. Na₂SO₄, evaporated in vacuo and dried in vacuo overnightto provide target product 5 (75 mg, 71%) as white solid. LC-MS [M+H]533.5 (C30H36N4O5+H, requires 533.6). Compound was used in nextsynthetic step without additional purification.

Synthesis ofN—[(S)-2-(2-dimethylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(98-2)

Reagent MW Eq. mg/mL mmol Compound 5 532.6 1.0 75 mg 0.14 NH₂OH × HCl69.49 6.0 58 mg 0.84 NaOtBu 96.11 8.0 108 mg  1.12 MeOH 1.5 mL THF 1.5mL

Compound was prepared using the same procedure as for synthesis ofcompound (98-1). LC-MS [M+H] 534.4 (C29H35N5O5+H, requires 534.6).

Retention Scale Yield Yield Purity [M + time Compound (mmol)* (mg) (%)*(%) H] (min)* 98-2 0.2 25.8 20% 100 534.4 3.51 *Based on the amount ofcompound 3. **HPLC-MS Method B

Synthesis of (S)-3-acetylamino-2- {4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl esterster (7)

To the mixture of compound 3 di-hydrochloride (104 mg, 0.20 mmol) andacetic anhydride (36 μl, 0.38 mmol) in DMF (0.5 mL) and CHCl₃ (1.5 mL)was added DIEA (210 μl, 1.2 mmol). Reaction mixture was stirred atambient temperature overnight. Solvent was concentrated in vacuo.Residue was dissolved in EtOAc (50 mL), washed with 5% solution ofNaHCO₃ (30 mL) and brine (30 mL). Organic phase was dried over anh.Na₂SO₄, evaporated in vacuo and dried in vacuo overnight to providetarget product 7 (70 mg, 71%) as white solid. LC-MS [M+H] 490.3(C28H31N3O5+H, requires 490.5). Compound was used in next synthetic stepwithout additional purification.

Synthesis ofN—((S)-2-acetylamino-1-hydroxycarbamoyl-ethyl)-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(98-3)

Reagent MW Eq. mg/mL mmol Compound 7 489.6.6 1.0 70 mg 0.14 NH₂OH × HCl69.49 6.0 58 mg 0.84 NaOtBu 96.11 8.0 108 mg  1.12 MeOH 1.5 mL THF 1.5mL

Compound was prepared using the same procedure as for synthesis ofcompound (98-1). LC-MS [M+H] 491.2 (C27H30N4O5+H, requires 491.5).

Retention Scale Yield Yield Purity [M + time Compound (mmol)* (mg) (%)*(%) H] (min)** 98-3 0.2 22.4 18.6% 100 491.2 3.41 *Based on the amountof compound 3. **HPLC-MS Method B

Example 994-[(E)-4-(4-{[Ethyl-(2-methoxy-ethyl)-amino]-methyl}-phenyl)-but-3-en-1-ynyl]-N—((S)-1-hydroxycarbamoyl-2-methylamino-ethyl)-benzamide(99-1)

Synthesis of (S)-3-hydroxy-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (2)

Reagent MW Eq. mg/ml mmol Compound 1 (Ach-Tr-Db- 347.41 1.0 139 mg 0.40(Morpholine)-OH H-(S)-Ser-OMe × HCl 155.58 1.2 75 mg 0.48 BOP 442.281.25 223 mg 0.50 DIEA 129.24 3.0 210 μl 1.20 DMF 4 ml

The product 2 (179 mg, 100%) as white solid was prepared using the BOPcoupling described in Example 96 for compound 2. LC-MS [M+H] 449.4(C26H28N2O5+H, requires 449.51). Compound was used in next syntheticstep without additional purification.

Synthesis of(S)-3-methylamino-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (3)

A solution of compound 2 (179 mg, 0.4 mmol) and DIEA (140 μl, 0.8 mmol)in CH₂Cl₂ (5 ml) was cooled to 0° C. followed by the addition of MsCl(37 μl, 0.48 mmol). Reaction mixture was maintained at 0° C. for 30 minfollowed by the addition of 2 M solution of methylamine in THF (2 ml, 4mmol). Temperature of the reaction mixture was raised to ambient.Reaction mixture was maintained at ambient temperature. After completion(in 30 min) solvent was removed in vacuo. Residue was dissolved in EtOAc(50 ml), washed with water (30 ml×2), brine (50 ml) and dried over anh.Na₂SO₄. Solvent was evaporated in vacuo and dried in vacuo overnight toprovide compound 3 (120 mg, 65%) as light yellow solid. LC-MS [M+H]462.4 (C27H31N3O4+H, requires 462.5).

Synthesis of 4- [(E)-4-(4-{[ethyl-(2-methoxy-ethyl)-amino]-methyl}-phenyl)-but-3-en-1-ynyl]-N—((S)-1-hydroxycarbamoyl-2-methylamino-ethyl)-benzamide(99-1)

Reagent MW Eq. mg/ml mmol Compound 3 461.55 1.0 120 mg 0.26 NH₂OH × HCl69.49 6.0 110 mg 1.56 NaOtBu 96.11 8.0 200 mg 2.1 MeOH 2 ml THF 2 ml

The product 99-1 (11.2 mg) as white solid was prepared by using the sameprocedure as for synthesis of compound (98-1). LC-MS [M+H] 463.3(C26H30N4O4+H, requires 463.5).

Retention Scale Yield Yield Purity [M + time Compound (mmol)* (mg) (%)*(%) H] (min)** 99-1 0.4 11.2 4.1 92.2 463.3 3.48 *Based on the amount ofcarboxylic acid used in the coupling reaction **HPLC-MS Method B

Example 100N—[(S)-2-(2-Cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(100-1)

Synthesis of (S)-3-tert-butoxycarbonylamino-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (2)

Reagent MW Eq. mg/mL mmol Compound 1 (Ach-Tr-Db- 347.41 1.0 139 mg 0.40(Morpholine)-OH N-Boc-(S)-DAP-OMe × HCl 254.71 1.2 122 mg 0.48 BOP442.28 1.25 223 mg 0.50 DIEA 129.24 3.0 210 μl 1.2 DMF 4 mL

The product 2 (207 mg, 94%) as yellow solid was prepared using the BOPcoupling described in Example 96 as for the synthesis of the compound 2.LC-MS [M+H] 548.4 (C31H37N3O6+H, requires 548.6). Compound was used innext synthetic step without additional purification.

Synthesis of(S)-3-amino-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 2 547.64 1.0 207 mg 0.38 4NHCl/dioxane 21 2 mL 8 Dioxane/MeOH (1:1) 2 mL

The product 3 (208 mg, 100%) as white solid was made using the GeneralMethod for Boc deprotection. LC-MS [M+H] 448.4 (C₂₆H₂₉N₃O₄+H, requires448.5). Compound was used in next synthetic step without additionalpurification.

Synthesis of(S)-3-(2-bromo-acetylamino)-2-[4-((3E,5E,7E)-5,8-dimethyl-9-morpholin-4-yl-nona-3,5,7-trien-1-ynyl)-benzoylamino]-propionicacid methyl ester (4)

To the mixture of compound 3 di-hydrochloride (207 mg, 0.40 mmol) andDIEA (180 μl) in CH₂Cl₂ (4 mL) was added bromoacetyl bromide (35 μl, 0.4mmol) at 0° C. Reaction mixture was allowed to warm to ambienttemperature and stirred for 30 min. Solvent was evaporated in vacuo.Residue was dissolved in EtOAc (50 mL), washed with 5% solution ofNaHCO₃ (50 mL) and brine (50 mL×2). Organic phase was dried over anh.Na₂SO₄, evaporated in vacuo and dried in vacuo overnight to providetarget product 4 (146 mg, 64%) as white solid. LC-MS [M+H] 570.1(C28H30BrN3O5+H, requires 569.5). Compound was used in next syntheticstep without additional purification.

Synthesis of (S)-3-(2-cyclopropylamino-acetylamino)-2-{4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (5)

To a solution of compound 4 (146 mg, 0.26 mmol) in CH₂Cl₂ (4 mL) wasadded cyclopropylamine (36 μl, 0.52 mmol) at 0° C. Reaction mixture wasmaintained at 0° C. for 10 min, then mixture was allowed to warm toambient temperature and stirred overnight. After completion the solventwas removed in vacuo. Residue was dissolved in EtOAc (60 mL), washedwith water (50 mL×2), brine (50 mL), dried over anh. Na₂SO₄ andconcentrated in vacuum. Residue was dissolved in DMSO (800 μl) andsubjected to HPLC purification. [YMC-Pack ODS-AC-18 column (30×100 mm);flow rate=40 mL/min; mobile phase A: 100% water, 0.1% TFA; mobile phaseB: 100% ACN, 0.1% TFA; gradient elution from 10% B to 50% B in 50 min.,detection 254 nm]. Fractions containing the desired product werecombined and evaporated in vacuo. Residue was dissolved in i-PrOH (10mL), evaporated in vacuum and dried in vacuo overnight to providedi-trifluoroacetate salt of the target product 5 (70 mg, 36%) as whitesolid. LC-MS [M+H] 545.0 (C₃₁H₃₆N₄O₅+H, requires 545.6).

Synthesis ofN—[(S)-2-(2-cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-3-en-1-ynyl]-benzamide(100-1)

Reagent MW Eq. mg/mL mmol Compound 5 × 2 TFA 738.6 1.0 70 mg 0.095 NH₂OH× HCl 69.49 6.0 40 mg 0.57 NaOtBu 96.11 8.0 73 mg 0.76 MeOH 1 mL THF 1mL

The product 100-1 (18.4 mg) as white solid was prepared using the sameprocedure as for synthesis of compound (98-1). LC-MS [M+H] 546.6(C30H35N5O5+H, requires 546.6).

Retention Scale Yield Yield Purity [M + time Compound (mmol)* (mg) (%)*(%) H] (min)** 100-1 0.4 18.4 6.0% 95 546.6 3.68 *Based on the amount ofcarboxylic acid used in the coupling reaction **HPLC-MS Method B

Example 101 Synthesis of(E)-4-[4-(4-Hydroxymethylphenyl)but-3-en-1-ynyl]benzoic acid methylester (1-2)

Step 1: 4-Hydroxymethylbenzaldehyde (2)

A suspension of terephthaldehyde (200 g, 1.492 mol, 1.0 eq.) in THF(1500 mL) was cooled to 0° C. NaBH₄ (16.94 g, 0.448 mol, 0.3 eq.) wasadded in one portion. The mixture was stirred at 1020° C. overnight. 800mL 1 N HCl was added and the mixture was stirred for 20 min and turnedclear. Most THF was removed in vacuum. Then, the residue was extractedby EtOAc (500 mL×3). The combined organic layer was washed with water(2×400 mL), brine (400 mL) and dried (MgSO₄), filtered and concentratedto give the compound 2 (200 g, 98%) which was used directly in next stepwithout purification.

Step 2: 4-(tert-Butyldimethylsilanyloxymethyl)benzaldehyde (3)

A solution of compound 2 (200 g, 1.47 mol, 1.0 eq.), TBSCl (220 g, 1.47mol, 1.0 eq.) and imidazole (200 g, 2.94 mol, 2.0 eq.) in DMF (1000 mL)was heated at 45° C. for 2 hrs. The mixture was concentrated to removemost DMF and DCM (2000 mL) was added. The mixture was washed with water(500 mL×2), brine (500 mL) and dried (MgSO₄), filtered and concentratedunder reduced pressure. The residue was purified by chromatography onsilica gel (0 to 5% EtOAc/PE) to give the compound 3 (130 g, 35.4%).¹H-NMR (300 MHz, CDCl₃): in ppm, 0.10 (s, 6H), 0.94 (s, 9H), 4.80 (s,2H), 7.47 (d, J=8.7 Hz, 2H), 7.84 (d, J=8.1 Hz, 2H), 9.98 (s, 1H).

Step 3: (E)-tert-Butyl-[4-(2-iodo-vinyl)benzyloxy]dimethylsilane (4)

Chromium chloride anhydrous (29.3 g, 240 mmol, 6 eq) was added to THF(500 mL) under nitrogen. A solution of triiodomethane (31.4 g, 80 mmol,2 eq) in THF (200 mL) was added dropwise at 0˜10° C. under nitrogen.Then, a solution of compound 3 (10.0 g, 40 mmol, 1 eq) in THF (100 mL)was added dropwise. The mixture was stirred at 0˜10° C. for 2 hrs andthen at 10-20° C. for 2 more hours. 400 mL iced water was added and themixture was stirred for 20 min, extracted with EtOAc (500 mL×3). Thecombined organic phase was washed with 20% aq. Na₂S₂O₃ aqueous (500 mL),water (500 mL), brine (500 mL) and dried (MgSO₄), filtered andconcentrated under reduced pressure. The residue was purified bychromatography on silica gel (0 to 2% EtOAc/PE) to give the titlecompound 4 (9 g, 60%). ¹H-NMR (300 MHz, CDCl₃): in ppm, 0.09 (s, 6H),0.93 (s, 9H), 4.72 (s, 2H), 0.6.79 (d, J=15 Hz, 1H), 7.26 (s, 4H), 7.41(d, J=15 Hz, 1H).

Step 4: (E)-4-{4-[4-(tert-Butyldimethylsilanyloxymethyl)phenyl]but-3-en-1-ynyl}benzoicacid methyl ester (6)

To a mixture of compound 4 (1.02 g, 2.7 mmol, 1.0 eq), 5 (0.43 g, 2.7mmol, 1.0 eq), PdCl₂(PPh₃)₂ (190 mg, 0.27 mmol, 0.1 eq) and Et₃N (5 mL)in THF (50 mL) was added CuI (51 mg, 0.27 mmol, 0.1 eq) under nitrogen.The mixture was stirred at 10-20° C. for 12 hrs. The reaction mixturewas diluted with EtOAc (50 mL), filtered and concentrated. The residuewas dissolved into EtOAc (100 mL) and washed with water (100 mL) andbrine (50 mL), and dried (MgSO₄). After filtration and concentration,the residue was purified by chromatography on silica gel (0 to 2%EtOAc/PE) to give the title compound 6 (0.625 g, 57%). ¹H-NMR (300 MHz,CDCl₃): in ppm, 0.10 (s, 6H), 0.94 (s, 9H), 3.92 (s, 3H), 4.76 (s, 2H),6.36 (d, J=18 Hz, 1H), 7.08 (d, J=18 Hz, 1H), 7.31 (d, J=9.0 Hz, 2H),7.40 (d, J=9.0 Hz, 2H), 7.52 (d, J=8.5 Hz, 2H), 7.99 (d, J=8.5 Hz, 2H).

Step 5: (E)-4-[4-(4-Hydroxymethylphenyl)but-3-en-1-ynyl]benzoic acidmethyl ester (I-2)

To a solution of compound 6 (5.65 g, 13.9 mmol, 1 eq) in THF (50 mL) wasadded Et₃N-3HF (6.72 g, 21.7 mmol, 3 eq) at room temperature. Themixture was stirred at 10˜20° C. for 10 hrs. The mixture wasconcentrated to remove solvent and EtOAc (100 mL), NaHCO₃ (50 mL) wasadded. The mixture was filtered and the organic of filtrate wasseparated, washed with brine (2×100 mL) and dried by MgSO₄, filtered.The filtrate was concentrated to 10 mL and filtered. The combined filtercake was dried in vacuum to give the target compound I-2 (3.25 g, 80%).¹H-NMR (300 MHz, DMSO-d₆): in ppm, 3.86 (s, 3H), 4.51 (d, J=5.76 Hz,2H), 5.25 (t, J=5.76 Hz, 1H) 6.65 (d, J=16.5 Hz, 1H), 7.17 (d, J=16.2Hz, 1H), 7.33 (d, J=8.0 Hz, 2H), 7.55 (d, J=8.2 Hz, 2H), 7.61 (d, J=8.25Hz, 2H), 7.97 (d, J=8.52 Hz, 2H).

N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzamide(101-1)

Synthesis of 4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzoicacid (2)

Reagent MW Eq. g/mL mmol Compound 1 293.33 1.0 780 mg 2.63(Ach-Triple-Double(OH)—OMe) 1N NaOH 2.5 6.5 mL 6.57 THF 4 mL MeOH 4 mL

The compound 2 (710 mg, 90%) as white solid was made and separated usingthe General Method for basic hydrolysis.

Synthesis of (2S,3R)-3-hydroxy-2-{4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-butyricacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 2 × sodium salt 300.28 1.0 99 mg 0.33H-(S)-Thr-OMe × HCl 169.6 1.2 66 mg 0.39 BOP 442.28 1.3 184 mg 0.42 DIEA129.24 3.0 172 μl 0.99 DMF 2 mL

The compound 3 (120 mg, 92%) as off-white solid was prepared using theBOP coupling described in Example 96 as for the synthesis of thecompound 2. LC-MS [M+H] 394.3 (C23H23NO5+H, requires 394.4). Compoundwas used in next synthetic step without additional purification.

Synthesis of 4N-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzamide(101-1)

Reagent MW Eq. mg/mL mmol Compound 3 393.4 1.1 120 mg 0.30 NH₂OH × HCl69.49 6.5 135 mg 1.94 25% NaOMe/MeOH 8.8 600 μl 2.64 MeOH 4 mL THF 1.5mL

The target product 101-1 (20.7 mg) as white solid made using the GeneralMethod for hydroxamate formation. LC-MS [M+H] 395.2 (C22H22N2O5+H,requires 395.4).

Com- Scale Yield Yield Purity Retention pound (mmol)* (mg) (%)* (%) [M +H] time (min)** 101-1 0.33 20.7 16.0 90.0 395.2 5.31 *Based on theamount of carboxylic acid used in the coupling reaction **HPLC-MS MethodBEach of the following compounds was synthesized as described above.

Ret. Compound MH⁺ Time HPLC-MS # Structure (m/z) (min) Method 101-2*

380.5 4.56 B 101-3

394.2 4.65 B 101-4

395.3 5.36 B 101-5**

409.2 5.61 B 101-6*

421.1 2.52 A *Boc-protecting group from intermediate was removed by 4MHCl/dioxane. **Methyl ester of diMeSer was prepared according to generalprocedure.

Example 1024-[(E)-4-(4-Cyclopropylaminomethyl-phenyl)-but-3-en-1-ynyl]-N-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide(102-1)

Synthesis of4-[(E)-4-(4-cyclopropylaminomethyl-phenyl)-but-3-en-1-ynyl]-benzoic acidmethyl ester (2)

A solution of compound 1 (750 mg, 2.56 mmol) and DIEA (1.1 mL, 6.40mmol) in CH₂Cl₂ (15 mL) was cooled to 0° C. followed by the addition ofMSCl (0.22 mL, 2.82 mmol) dropwise over the period of 5 min. Reactionmixture was maintained at 0° C. for 10 min followed by the addition ofcyclopropylamine (0.89 mL, 12.8 mmol). Temperature of the reactionmixture was allowed to rise to ambient. Reaction mixture was maintainedat ambient temperature overnight. After completion solvent was removedin vacuo. Residue was dissolved in EtOAc (150 mL), washed with water(150 mL×2), brine (150 mL) and dried over anh. Na₂SO₄. Solvent wasevaporated in vacuo to provide compound 2 (848 mg, 100%) as light yellowoil. LC-MS [M+H] 332.2 (C22H21NO2+H, requires 332.41). Compound was usedin next synthetic step without additional purification.

Synthesis of 4-((E)-4-{4-[(tert-butoxycarbonyl-cyclopropyl-amino)-methyl]-phenyl}-but-3-en-1-ynyl)-benzoicacid (3)

Reagent MW Eq. g/mL mmol Compound 2 331.41 1.0 848 mg 2.56 1N aq NaOH3.1 8 mL 8.0 Di-tert-butyl dicarbonate 218.2 1.1 615 mg 2.82 THF 8 mLdioxane 5 mL

The compound 3 (795 mg, 75%) as white solid was made and separated usingthe General Method for Boc protection. LC-MS [M+H] 418.3 (C26H27NO4+H,requires 418.5).

Synthesis of (2S,3R)-2-[4-((E)-4-{4-[(tert-butoxycarbonyl-cyclopropyl-amino)-methyl]-phenyl}-but-3-en-1-ynyl)-benzoylamino]-3-hydroxy-butyricacid methyl ester (4)

Reagent MW Eq. mg/mL mmol Compound 3 417.5 1.0 155 mg 0.37 H-(S)-Thr-OMe× HCl 169.6 1.2 75 mg 0.44 BOP 442.28 1.3 212 mg 0.48 DIEA 129.24 3.0190 μl 1.10 DMF 2.5 mL

The product 4 (140 mg, 71%) as light yellow oil was prepared using theBOP coupling described in Example 96 as for the synthesis of thecompound 2. LC-MS [M+H] 533.4 (C31H36N2O6+H, requires 533.6). Compoundwas used in next synthetic step without additional purification.

Synthesis of(2S,3R)-2-{4-[(E)-4-(4-cyclopropylaminomethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-3-hydroxy-butyricacid methyl ester (5)

Reagent MW Eq. mg/mL mmol Compound 4 532.6 1.0 140 mg 0.26 4NHCl/dioxane 23.0 1.5 mL 6.0 dioxane 1.5 mL

The product 5 (121 mg, 100%) as white solid was made using the GeneralMethod for Boc deprotection. LC-MS [M+H] 433.4 (C26H28N2O4+H, requires433.5). Compound was used in next synthetic step without additionalpurification.

Synthesis of4-[(E)-4-(4-cyclopropylaminomethyl-phenyl)-but-3-en-1-ynyl]-N-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-benzamide(102-1)

Reagent MW Eq. mg/mL mmol Compound 5 × HCl 468.97 1.0 121 mg 0.26 NH₂OH× HCl 69.49 7.4 135 mg 1.94 25% NaOMe/MeOH 10.0 600 μl 2.64 MeOH 4 mLTHF 1.5 mL

The product 102-1 (57.6 mg) as white solid was made using the GeneralMethod for hydroxamate formation. LC-MS [M+H] 434.4 (C25H27N3O4+H,requires 434.5).

Com- Scale Yield Yield Purity Retention pound (mmol)* (mg) (%)* (%) [M +H] time (min)** 102-1 0.37 57.6 28.5 90.1 434.3 4.41 *Based on theamount of carboxylic acid used in the coupling reaction **HPLC-MS MethodBEach of the following compounds was synthesized as described above.

Ret. Compound MH⁺ Time HPLC-MS # Structure (m/z) (min) Method 102-2

433.2 3.86 B 102-3

419.6 3.69 B 102-4

434.3 4.44 B 102-5*

448.5 4.69 B 102-6

447.9 2.74 A *Methyl ester of di-Me-Ser was prepared according togeneral procedure. **Boc-protecting group before hydroxamate formationwas removed by 2N HCl/dioxane

Example 103N—[(S)-2-(2-Cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzamide(103-1)

Synthesis of(S)-3-tert-butoxycarbonylamino-2-{4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (2)

Reagent MW Eq. mg/mL mmol Compound 1 × sodium salt 300.28 1.0 120 mg0.40 H-(S)-DAP(Boc)-OMe × HCl 254.7 1.2 122 mg 0.48 BOP 442.28 1.3 230mg 0.52 DIEA 129.24 3.0 210 μl 1.2 DMF 3 mL

The compound 2 (191 mg, 100%) as off-white solid was prepared usingprocedure of the BOP coupling described in Example 96 as for thesynthesis of the compound 2. LC-MS [M+H] 479.3 (C27H30N2O6+H, requires479.5). Compound was used in next synthetic step without additionalpurification.

Synthesis of(S)-3-amino-2-{4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 2 478.5 1.0 191 mg 0.4 4N HCl/dioxane30 3 mL 12 Dioxane/MeOH (1:1) 3 mL

The product 3 (165 mg, 100%) as white solid was made using the GeneralMethod for Boc deprotection. LC-MS [M+H] 379.1 (C22H22N2O4+H, requires379.4).

Synthesis of ((S)-3-(2-bromo-acetylamino)-2-{4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (4)

Reagent MW Eq. mg/mL mmol Compound 3 × HCl 414.9 1.0 165 mg 0.40Bromoacetyl bromide 201.86 1.0 35 μl 0.40 DIEA 129.24 2.5 180 μl 1.0CH₂Cl₂ 4 mL DMF 1 mL

The target product 4 (199 mg, 100%), as yellow solid, was prepared usingthe method described in compound 4 of Example 100. LC-MS [M+H] 501.3(C24H23BrN2O5+H, requires 500.4). Compound was used in next syntheticstep without additional purification.

Synthesis of(S)-3-(2-cyclopropylamino-acetylamino)-2-{4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzoylamino}-propionicacid methyl ester (5)

Reagent MW Eq. g/mL mmol Compound 4 499.35 1.0 199 mg 0.40Cyclopropylamine 57.09 3.7 100 μl 1.48 CH₂Cl₂ 4 mL

The target compound 5 (124 mg, 65%) as solid was prepared using themethod described in compound 5 of Example 100. LC-MS [M+H] 476.3(C27H29N3O5+H, requires 476.6).

Synthesis ofN—[(S)-2-(2-cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-[(E)-4-(4-hydroxymethyl-phenyl)-but-3-en-1-ynyl]-benzamide(103-1)

Reagent MW Eq. mg/mL mmol Compound 5 475.6 1.0 124 mg 0.26 NH₂OH × HCl69.49 7.4 135 mg 1.94 25% NaOMe/MeOH 10.0 600 μl 2.64 MeOH 4 mL THF 1.5mL

The product 6 (12.2 mg) as white solid was made using the General Methodfor hydroxamate formation. LC-MS [M+H] 477.3 (C26H28N4O5+H, requires477.5).

Retention Scale Yield Yield Purity time Compound (mmol)* (mg) (%)* (%)[M + H] (min)** 103-1 0.4 12.2 5.2 98.9 477.3 4.8 *Based on the amountof carboxylic acid used in the coupling reaction **HPLC-MS Method B

Example 104 Synthesis of (E)-4-(4-Phenylbut-3-en-1-ynyl)benzoic acid(I-3)

(E)-(2-Iodovinyl)benzene (2)

Chromium chloride anhydrous (37.1 g, 302 mmol, 8 eq) was added to THF(400 mL) under nitrogen at room temperature. A solution oftriiodomethane (59.7 g, 151 mmol, 4 eq) in THF (500 mL) was addeddropwise at 0° C. under nitrogen. Then a solution of benzaldehyde 1 (4.0g, 37.7 mmol, 1 eq) in THF (100 mL) was added dropwise. The mixture wasstirred at 0° C. for 2 hrs and then at room temperature for 2 morehours. The mixture was poured into ice water and extracted with EtOAc(800 mL×2). The organic phase was washed with 20% aq. Na₂S₂O₃ (300 mL),water (800 mL), brine (300 mL) dried (MgSO₄), filtered and concentratedunder reduced pressure. The residue was triturated with EtOAc andhexanes. After filtration, the filtrates were concentrated to give thetitle compound 2 (7.2 g, 83%). ¹H NMR (300 MHz, CDCl₃): δ ppm, 6.83 (d,J=15.1 Hz, 1H), 7.29-7.32 (m, 5H), 7.43 (d, J=14.8 Hz, 1H).

(E)-4-(4-Phenylbut-3-en-1-ynyl)benzoic acid methyl ester (3)

To a mixture of compound 2 (3.77 g, 16.4 mmol, 1.0 eq), 4 (2.62 g, 16.4mmol, 1.0 eq), PdCl₂(PPh₃)₂ (0.58 mg, 0.82 mmol, 0.05 eq) and Et₃N (10mL) in THF (100 mL) was added CuI (0.32 g, 1.64 mmol, 0.1 eq) undernitrogen at room temperature. The mixture was stirred at roomtemperature for 12 hrs. The reaction mixture was diluted with EtOAc (50mL), filtered and concentrated. The residue was dissolved into EtOAc(300 mL) washed with water (300 mL) and brine (150 mL), and dried(MgSO₄). After filtration and concentration, the residue was purified bychromatography on silica gel (0 to 15% EtOAc/Hexanes) to give the titlecompound 3 (2.56 g, 60%). ¹H NMR (300 MHz, DMSO-d₆): δ ppm, 3.87 (s,3H), 6.70 (d, J=16.2 Hz, 1H), 7.19 (d, J=16.2 Hz, 1H), 7.35-7.43 (m,3H), 7.59-7.64 (m, 4H), 7.97 (d, J=8.22 Hz, 2H)

(E)-4-(4-Phenylbut-3-en-1-ynyl)benzoic acid (1-3)

The product I-3 (1.45 g, 61%) was made and separated using the GeneralMethod for basic hydrolysis. MS (m/z): [M−H]+=247, ¹H NMR (300 MHz,DMSO-d₆) δ ppm, 6.70 (d, J=16.2 Hz, 1H), 7.19 (d, J=16.2 Hz, 1H),7.32-7.42 (m, 3H), 7.58-7.61 (m, 4H), 7.95 (d, J=8.5 Hz, 2H).

N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-((E)-4-phenyl-but-3-en-1-ynyl)-benzamide(104-1)

Synthesis of(2S,3R)-3-hydroxy-2-[4-((E)-4-phenyl-but-3-en-1-ynyl)-benzoylamino]-butyricacid methyl ester (2)

Reagent MW Eq. mg/mL mmol Compound 1 (Ach-Tr-Db-OH) 248.28 1.0 85 mg0.34 H-(S)-Thr-OMe × HCl 169.6 1.1 63 mg 0.37 HATU 380.2 1.2 155 mg 0.41DIEA 129.24 3.0 178 μl 1.0 DMF 2 mL

The product 2 (123 mg, 100%) as white solid was prepared using theGeneral Method for HATU coupling. LC-MS [M+H] 364.7 (C22H21NO4+H,requires 364.42). Compound was used in next synthetic step withoutadditional purification.

Synthesis ofN-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-((E)-4-phenyl-but-3-en-1-ynyl)-benzamide(23)

Reagent MW Eq. mg/mL mmol Compound 2 363.4 1.0 123 mg 0.34 NH₂OH × HCl69.49 8.0 188 mg 2.7 25% NaOMe/MeOH 10.0 777 μl 3.4 MeOH 4 mL THF 1.5 mL

The product 104-1 (58.7 mg, 48%) as white solid was made using theGeneral Method for hydroxamate formation. LC-MS [M+H] 365.4(C21H20N2O4+H, requires 365.4).

Retention Scale Yield Yield Purity time Compound (mmol)* (mg) (%)* (%)[M + H] (min)** 104-1 0.34 58.7 48 98.0 365.4 7.28 *Based on the amountof carboxylic acid used in the coupling reaction **HPLC-MS Method BEach of the following compounds was synthesized as described above.

HPLC- Compound MH⁺ Ret. Time MS # Structure (m/z) (min) Method 104-2

350.7 6.38 B 104-3*

379.7 7.58 B 104-4

364.7 6.46 B 104-5**

378.5 4.55 A *Methyl ester of diMeSer was prepared according to generalprocedure. **Boc-protecting group before hydroxamate formation wasremoved by 4N HCl/dioxane

Example 105N—[(S)-2-(2-Cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-((E)-4-phenyl-but-3-en-1-ynyl)-benzamide(105-1)N—{(S)-1-Hydroxycarbamoyl-2-[2-((S)-1-phenyl-ethylamino)-acetylamino]-ethyl}-4-((E)-4-phenyl-but-3-en-1-ynyl)-benzamide(105-2)

Synthesis of(S)-3-tert-butoxycarbonylamino-2-[4-((E)-4-phenyl-but-3-en-1-ynyl)-benzoylamino]-propionicacid methyl ester (2)

Reagent MW Eq. mg/mL mmol Compound 1 (Ach-Tr-Db-OH) 248.3 1.0 189 mg0.76 H-(S)-DAP(Boc)-OMe × HCl 254.7 1.2 232 mg 0.91 HATU 380.2 1.2 346mg 0.91 DIEA 129.2 3.0 397 μl 2.28 DMF 4 mL

The product 2 (328 mg, 100%) as off-white solid was prepared using theGeneral Method for HATU coupling. LC-MS [M+H] 433.8 (C26H28N2O4+H,requires 433.52). Compound was used in next synthetic step withoutadditional purification.

Synthesis of((S)-3-amino-2-[4-((E)-4-phenyl-but-3-en-1-ynyl)-benzoylamino]-propionicacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 2 432.5 1.0 328 mg 0.76 4NHCl/dioxane 31.6  6 mL 24.0

The product 3 (292 mg, 100%) as white solid was made using the GeneralMethod for Boc deprotection. LC-MS [M+H] 349.7 (C21H20N2O3+H, requires349.41).

Synthesis of(S)-3-[2-(tert-butoxycarbonyl-cyclopropyl-amino)-acetylamino]-2-[4-((E)-4-phenyl-but-3-en-1-ynyl)-benzoylamino]-propionicacid methyl ester (4)

Reagent MW Eq. mg/mL mmol Compound 3 × HCl 384.9 1.0 146 mg 0.38N-Boc-N-Cyclopropyl-Gly-OH 215.25 1.1  90 mg 0.42 HATU 380.2 1.2 175 mg0.46 DIEA 129.2 3.0 198 μl 1.14 DMF  3 mL

The product 4 (207 mg, 100%) as light yellow oil was prepared using theGeneral Method for HATU coupling. LC-MS [M+H] 546.4 (C31H35N3O6+H,requires 546.64). Compound was used in next synthetic step withoutadditional purification.

Synthesis of(S)-3-(2-cyclopropylamino-acetylamino)-2-[4-((E)-4-phenyl-but-3-en-1-ynyl)-benzoylamino]-propionicacid methyl ester (5)

Reagent MW Eq. mg/mL mmol Compound 4 545.6 1.0 207 mg 0.38 4NHCl/dioxane 31.6  3 mL 12.0

The product 5 (183 mg, 100%) as white solid was made using the GeneralMethod for Boc deprotection. LC-MS [M+H] 446.3 (C₂₆H₂₇N₃O₄+H, requires446.52).

Synthesis ofN—[(S)-2-(2-cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-((E)-4-phenyl-but-3-en-1-ynyl)-benzamide(105-1)

Reagent MW Eq. mg/mL mmol Compound 5 × HCl 481.97 1.0 183 mg 0.38 NH₂OH× HCl 69.49 8.0 211 mg 3.0 25% NaOMe/MeOH 10.0  868 μl 3.8 MeOH   4 mLTHF 1.5 mL

The product 105-1 (21.7 mg, 10%) as white solid was made using theGeneral Method for hydroxamate formation. LC-MS [M+H] 447.3(C25H26N4O4+H, requires 447.52).

Retention Scale Yield Yield Purity time Compound (mmol)* (mg) (%)* (%)[M + H] (min)** 105-1 0.38 21.7 10 99.0 447.3 6.49 *Based on the amountof carboxylic acid used in the coupling reaction **HPLC-MS Method B

Synthesis of(S)-3-(2-bromo-acetylamino)-2-[4-((E)-4-phenyl-but-3-en-1-ynyl)-benzoylamino]-propionicacid methyl ester (7)

Reagent MW Eq. mg/mL mmol Compound 3 × HCl 384.9 1.0 146 mg 0.38Bromoacetyl bromide 201.86  1.08  36 μl 0.41 DIEA 129.24 2.6 174 μl 1.0CH₂Cl₂ 3 mL

The target product 7 (150 mg, 85%), as yellow solid, was prepared usingthe method described in compound 4 of Example 19. LC-MS [M+H] 470.7(C23H21BrN2O4+H, requires 470.34).

Synthesis of(S)-2-[4-((E)-4-phenyl-but-3-en-1-ynyl)-benzoylamino]-3-[2-((S)-1-phenyl-ethylamino)-acetylamino]-propionicacid methyl ester (8)

Reagent MW Eq. mg/mL mmol Compound 7 469.3 1.0 150 mg 0.32(S)-(−)-alpha-Methylbenzylamine 121.18 3.0 124 μl 0.96 CH₂Cl₂  3 mL

The target compound 8 (155 mg, 95%) as a solid, was prepared using themethod described in compound 5 of Example 19. LC-MS [M+H] 510.3(C31H31N3O4+H, requires 510.61).

N—{(S)-1-Hydroxycarbamoyl-2-[2-((S)-1-phenyl-ethylamino)-acetylamino]-ethyl}-4-((E)-4-phenyl-but-3-en-1-ynyl)-benzamide(105-2)

Reagent MW Eq. mg/mL mmol Compound 8 509.61 1.0 155 mg 0.30 NH₂OH × HCl69.49 8.0 167 mg 2.4 25% NaOMe/MeOH 10.0  685 μl 3.0 MeOH   3 mL THF 1.2mL

The product 105-2 (41.7 mg) as white solid was made using the GeneralMethod for hydroxamate formation. LC-MS [M+H] 511.8 (C30H30N4O4+H,requires 511.61).

Com- Scale Yield Yield Purity Retention pound (mmol)* (mg) (%)* (%) [M +H] time (min)** 105-2 0.38 41.7 17.5 97.3 511.8 7.38 *Based on theamount of carboxylic acid used in the coupling reaction **HPLC-MS MethodBEach of the following compounds was synthesized as described above.

Ret. Compound MH⁺ Time HPLC-MS # Structure (m/z) (min) Method 105-3

460.4 7.79 B 105-4

511.8 7.37 B 105-5

525.2 7.56 B

Example 106 Methyl 4-[(3E)-4-chlorobut-3-en-1-yn-1-yl]benzoate

Methyl 4-[(3E)-4-chlorobut-3-en-1-yn-1-yl]benzoate

Methyl 4-ethynylbenzoate (2.0 g, 12.5 mmol, 1 eq) was dissolved in 200mL anhydrous THF under nitrogen. n-Butylamine (2.5 mL, 25.3 mmol, 2 eq)was added followed by trans-1,2-dichloroethylene (1.95 mL, 25.3 mmol, 2eq). Bis(triphenylphosphine)palladium(II) dichloride (0.44 g, 0.63 mmol,0.05 eq) was added as a solid, and the reaction mixture was stirred for20-30 minutes under nitrogen. Copper iodide (0.26 g, 1.4 mmol, 0.11 eq)was added as a solid to the clear amber solution, which became very darkwithin ten minutes. Stirring under nitrogen at room temperature wascontinued overnight. The reaction mixture was filtered through Celiteand THF passed through the filtercake until the filtrate was colorless.The volatiles were removed via rotary and the dark red-brown residuepartitioned between ethyl acetate and water. The layers were separatedand the organic layer washed with brine, dried over Na₂SO₄, and strippedto give the product as a dark brown solid. The crude material waspurified by chromatography on silica gel using 2.5% ethyl acetate inhexanes as eluent. Yield: 1.55 g, 56%. TLC R_(f)=0.46 (1:9 ethylacetate:hexanes). APCI(−) m/z=219 amu as a minor signal. ¹H NMR (CDCl₃,300 MHz) δ ppm: 7.99 (d, J=8.5 Hz, 2H), 7.48 (d, J=8.5 Hz, 2H), 6.67 (d,J=13.8 Hz, 2H), 6.16 (d, J=13.5 Hz, 2H), 3.92 (s, 3H).

Methyl 4-[(3E)-4-pyridin-4-ylbut-3-en-1-yn-1-yl]benzoate

Methyl 4-[(3E)-4-pyridin-4-ylbut-3-en-1-yn-1-yl]benzoate

Methyl 4- [(3E)-4-chlorobut-3-en-1-yn-1-yl]benzoate (47 mg, 0.21 mmol, 1eq), 4-pyridylboronic acid (28 mg, 0.23 mmol, 1.1 eq) andtetra-n-butylammonium bromide (58 mg, 0.21 mmol, 1 eq) were placed in aflask and dissolved in 5 mL toluene. Aqueous K₃PO₄ (3 M, 0.20 mL, 0.60mmol, 2.8 eq) was added, followed bytetrakis(triphenylphosphine)palladium(0) (18 mg, 0.016 mmol, 0.08 eq).The reaction mixture was heated at reflux for three hours, then stirredat room temperature overnight. TLC in 1:9 ethyl acetate:hexanesconfirmed the absence of starting material. The reaction mixture wasdiluted with ethyl acetate and washed sequentially with water and brine,then the volatiles were removed via rotary. The residue was plated ontosilica and eluted with 33% ethyl acetate/hexanes to give the product asa yellow solid. Yield: 26 mg (46%). TLC R_(f)=0.20 (40% ethylacetate/hexanes). APCI(+) m/z=246 amu. ¹H NMR (CDCl₃, 300 MHz) δ ppm:8.60 (d, J=6.1 Hz, 2H), 8.02 (d, J=8.3 Hz, 2H), 7.54 (d, J=8.5 Hz, 2H),7.28 (d, J=6.3 Hz, 2H), 6.99 (d, J=16.2 Hz, 2H), 6.59 (d, J=16.2 Hz,2H), 3.93 (s, 3H).

N-((1S,2R)-2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-((E)-4-pyridin-4-yl-but-3-en-1-ynyl)-benzamide(106-1)

Synthesis of(2S,3R)-3-hydroxy-2-[4-((E)-4-pyridin-4-yl-but-3-en-1-ynyl)-benzoylamino]-butyricacid methyl ester (2)

Reagent MW Eq. mg/mL mmol Compound 1 × sodium salt 271.25 1.0 109 mg 0.4H—(S)-Thr-OMe × HCl 169.6 1.2  82 mg 0.48 HATU 380.2 1.2 183 mg 0.48DIEA 129.24 3.0 209 μl 1.2 DMF  2.5 mL

The compound 2 (146 mg, 100%) as light yellow solid was prepared usingthe General Method for HATU coupling. LC-MS [M+H] 365.7 (C21H20N2O4+H,requires 365.4). Compound was used in next synthetic step withoutadditional purification.

Synthesis ofN-((1S,2R)-2-hydroxy-1-hydroxycarbamoyl-propyl)-4-((E)-4-pyridin-4-yl-but-3-en-1-ynyl)-benzamide(106-1)

Reagent MW Eq. mg/mL mmol Compound 2 364.4 1.0 146 mg 0.4 NH₂OH × HCl69.49 8.0 222 mg 3.2 25% NaOMe/MeOH 10.0  914 μl 4.0 MeOH 5 mL THF 2 mL

The product 106-1 (52 mg) as off-white solid was made using the GeneralMethod for hydroxamate formation. LC-MS [M+H] 366.8 (C20H19N3O4+H,requires 366.4).

Retention Com- Scale Yield Yield Purity time pound (mmol)* (mg) (%)* (%)[M + H] (min)** 106-1 0.4 52 mg 27.2% 99.2 366.8 2.30 *Based on theamount of carboxylic acid used in the coupling reaction **HPLC-MS MethodAEach of the following compounds was synthesized as described above.

Ret. Compound MH⁺ Time HPLC-MS # Structure (m/z) (min) Method 106-2*

380.6 3.22 B 106-3**

365.8 1.97 B 106-4

365.9 1.98 A 106-5

380.8 3.22 B 106-6

382.8 4.46 A 106-7

397.7 5.14 A *Methyl ester of di-Me-Ser was prepared according togeneral procedure. **Fmoc-protecting group before hydroxamate formationwas removed by 20% piperidine/EtOAc.

Example 1074-((E)-4-Cyclopropyl-but-3-en-1-ynyl)-N—((S)-2-hydroxy-1-hydroxycarbamoyl-2-methyl-propyl)-benzamide(107-1)

Synthesis of ((E)-2-Iodo-vinyl)-cyclopropane (2)

The chromium (II) chloride (15.0 g, 122.1 mmol) was placed in a flask inthe glove box with a stir bar and capped with a septum. With goodstirring, the THF (anh., 120 mL) was added quickly at ambienttemperature. After stirring for 15-30 min under nitrogen, the flask wasimmersed in an ice bath for 30 min. To the cold suspension was addeddropwise a solution of iodoform (11.42 g, 29.0 mmol) in THF (anh., 75mL). After 5 min, the solution of cyclopropylaldehyde 1 (2.43 mL, 30.5mmol) in THF (anh., 60 mL) was added neat dropwise to the vigorouslystirred suspension. After stirring at 0° C. for 2 h, the reaction wasallowed to warm to room temperature overnight. The reaction mixture waspoured into and ice water and extracted with pentane (300 mL×2). Theorganic phase was washed with 20% aq. Na₂S₂O₃ (150 mL), water (300 mL),brine (300 mL), then dried over MgSO4 (anh.), filtered and pentane wasconcentrated in vacuo at 220 mbar to minimum volume (20 mL). The residuewas used in the next synthetic step without additional purification(caution: cyclopropyl vinyl iodide is volatile material).

Synthesis of 4-((E)-4-cyclopropyl-but-3-en-1-ynyl)-benzoic acid methylester (3)

A solution of 4-ethynyl-benzoic acid methyl ester (1.76 g, 11.0 mmol),diisopropylamine (4.7 mL, 33 mmol), the catalysts PdCl₂(dppf)₂ (302 mg,0.37 mmol) and the copper (I) iodide (46 mg, 0.24 mmol) in THF (anh., 30mL) was purged for 10 min with dry nitrogen. Then, this mixture wasadded to a pentane solution of compound 1 (13.5 mL, C=˜200 mg/mL, ˜2.7g, 14 mmol). Reaction mixture was heated at 80° C. for 20 min and cooledto ambient temperature. The reaction mixture was then dissolved in EtOAc(200 mL), washed with water (150 mL), 0.5 N HCl (150 mL) and brine (150mL×2). Organic phase was dried over anh. Na2SO4, evaporated in vacuo.Residue was subjected to flash chromatography on CombiFlash® Companionunit equipped with RediSep® flash column (normal phase, 35-60 micronaverage particle size silica gel, 40 g, Teledyne Isco); flow rate=40mL/min; mobile phase A: hexane; mobile phase B: EtOAc; gradient 0-30% Bin 60 min. Fractions containing the desired material were combined andconcentrated in vacuo to provide target material 3 (1 g, 40%) as yellowoil. LC-MS [M+H] 227.7 (C15H14O2+H, requires 227.29).

Synthesis of 4-((E)-4-cyclopropyl-but-3-en-1-ynyl)-benzoic acid (4)

Reagent MW Eq. g/mL mmol Compound 3 226.28 1.0 1.0 g 4.41 1N NaOH 4.0 18 mL 18.0 THF/MeOH (1:1)  18 mL

The compound 4 (800 mg, 85%) as a white solid was made and separatedusing the General Method for basic hydrolysis. LC-MS [M+H] 213.5(C14H12O2+H, requires 213.3). Compound was used in next synthetic stepwithout additional purification. Check of a sample by LC-MS showed amajor product peak (Rt=5.10 min, E-isomer) with ˜12% of the cis-isomer(Rt=4.96 min) (HPLC-MS Method A).

Synthesis of(S)-2-[4-((E)-4-cyclopropyl-but-3-en-1-ynyl)-benzoylamino]-3-hydroxy-3-methyl-butyricacid methyl ester (5)

Reagent MW Eq. mg/mL mmol Compound 4 212.25 1.0  85 mg 0.4H—(S)-diMe-Ser-OMe × HCl* 169.6 1.13  76 mg 0.45 HATU 380.2 1.2 183 mg0.48 DIEA 129.24 3.0 209 μl 1.2 DMF  2 mL *Methyl ester of di-Me-Ser wasprepared according to general procedure.

The compound 5 (130 mg, 96%) as white solid was prepared using theGeneral Method for HATU coupling. LC-MS [M+H] 342.8 (C20H23N04+H,requires 342.4).

Synthesis of4-((E)-4-cyclopropyl-but-3-en-1-ynyl)-N—((S)-2-hydroxy-1-hydroxycarbamoyl-2-methyl-propyl)-benzamide(107-1)

Isopropyl alcohol (2 mL) was added to triple-double ester 5 (130 mg,0.38 mmol) and the mixture was cooled in an ice bath for 5 min. NH₂OH(50% aq, 2 mL, 32.8 mmol) was added to the mixture. After 5 min, the icebath was removed and the reaction mixture was stirred until reaction wascomplete (˜16 h, as determined by LC-MS analysis). Solvent volume wasreduced by half using a nitrogen stream and water (8 mL) was added. Thesuspension was thoroughly agitated (vibro mixer and sonication),centrifuged and the supernatant was discarded. Water (8 mL) was added tothe solid and the suspension was thoroughly agitated, centrifuged andthe supernatant was discarded. Solid was dissolved in DMSO (600 μl) andsubjected to HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 mL/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 8% B to 28% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and lyophilized to provide the target product (66.3 mg) aswhite solid. LC-MS [M+H] 343.7 (C19H22N2O4+H, requires 343.4).

Com- Scale Yield Yield Purity [M + Retention pound (mmol)* (mg) (%)* (%)H] time (min)** 107-1 0.4 66.3 mg 48.4% 97.5 343.7 6.41 *Based on theamount of carboxylic acid used in the coupling reaction **HPLC-MS MethodBEach of the following compounds was synthesized as described above.

Ret. Compound MH⁺ Time HPLC-MS # Structure (m/z) (min) Method 107-2

328.7 3.86 A 107-3

329.8 4.32 B 107-4*

342.7 5.59 B 107-5*

330.7 6.01 A 107-6*

344.8 6.14 B *Boc-protecting group before hydroxamate formation wasremoved by 2N HCl/dioxane **Desired scaffold for this product wasprepared starting from isobutyraldehyde

Example 108 Synthesis of (E)-4-(hex-3-en-1-ynyl) benzoic acid (IC-10)

(E)-1-iodobut-1-ene (2)

To but-1-yne (1.5 g, 27.78 mmol) in a sealable tube at 0° C. was addedDIBAL-H in hexane (1M; 28 mL, 28 mmol). The tube was sealed and allowedto stand for 30 min at R.T. before being heated to 55-60° C. for 4 h.After cooling down, the reaction mixture was evaporated in vacuo (usinga vacuum manifold) and the residue was dissolved in THF (15 mL) at 0° C.The resultant solution was cooled to −78° C. and I₂ (8.47 g, 33.3 mmol)in THF (30 mL) was added dropwise. The mixture was allowed to warm to 0°C. And transferred via cannula to a stirred mixture of 1 M HCl (70 mL)and Et₂O (40 mL). the layers were separated and the aqueous layer wasextracted with saturated aqueous sodium thioslufate (40 mL), 1M NaOH (40mL), distilled water (40 mL) and brine (40 ml), dried over Na₂SO₄,filtered, and rotary evaporated at 0° C. to give compound 2 as a brownliquid and to next step without purification.

(E)-Methyl 4-(hex-3-en-1-ynyl) benzoate (4)

Reagent MW Eq. mmol g, mL Compound 2 182.00 1.0 16.2 2.94 g Methyl4-ethynylbenzoate 160.17 1.0 16.2 2.58 g PdCl₂(PPh₃)₂ 701.90 0.1 1.621.13 g CuI 190.45 0.1 1.62 0.216 g TEA 101.19 13.0 220 30 mL THF 150 mL

The compound 4 (1.8 g, 52%) as a light yellow liquid was prepared byusing the method described in compound 5 of Example 96. ¹H NMR (500 MHz,CDCl₃): δ ppm, 1.06 (t, J=15 Hz, 3H), 2.2 (m, 2H), 3.91 (s, 3H), 5.70(d, J=15.5 Hz, 1H), 6.35 (m, 1H), 7.46 (d, J=8 Hz, 2H), 7.97 (d, J=8.5Hz, 2H).

(E)-4-(hex-3-en-1-ynyl) benzoic acid (IC-10)

Reagent MW Eq. mmol g, mL Compound 4 214.26 1.0 8.41 1.8 g NaOH(2Nsolution)   9 mL THF   9 mL Methanol   9 mL

The compound IC-10 (1.5 g, 89%) as a white solid was made and separatedusing the General Method for basic hydrolysis. ¹H NMR (500 MHz,DMSO-d₆): δ ppm, 1.0 (t, J=15 Hz, 3H), 2.18 (m, 2H), 5.83 (d, J=16 Hz,1H), 6.37 (m, 1H), 7.53 (d, J=8.5 Hz, 2H), 7.92 (d, J=8.5 Hz, 2H), 13.12(s, 1H)

Synthesis of (E)-4-(4-(1-methyl-1H-pyrazol-4-yl)but-3-en-1-ynyl) benzoicacid (IC-11)

(E)-4-(2-iodovinyl)-1-methyl-1H-pyrazole (2)

Reagent MW Eq. mmol g, mL Compound 1 110.11 1.0 8.2 900 mg Chromiumchloride 122.90 6.0 49.2 5.555 g triiodomethane 393.73 4.0 32.8 12.914 gTHF 185 mL

The target compound 2 (635 mg, 33%) was prepared by using the methoddescribed in compound 5 of Example 96. [M+1]: 234.9. ¹H-NMR: (400 MHz,DMSO-d₆): δ 3.87 (s, 3H), 6.43 (d, J=14.8 Hz, 1H), 7.21 (s, J=14.8 Hz,1H), 7.34 (s, 1H), 7.49 (s, 1H).

(E)-methyl 4-(4-(1-methyl-1H-pyrazol-4-yl)but-3-en-1-ynyl)benzoate (4)

To a mixture of compound (E)-4-(2-iodovinyl)-1-methyl-1H-pyrazole (635mg, 2.7 mmol, 1.0 equiv), methyl 4-ethynylbenzoate (434 g, 2.7 mmol, 1.0equiv), PdCl₂(PPh₃)₂ (190 mg, 0.3 mmol, 0.1 equiv) and (i-Pr)₂NH (546mg, 5.4 mmol, 2 equiv) in THF (100 mL) was added CuI (51 mg, 0.3 mmol,0.1 equiv) under nitrogen at room temperature. The mixture was stirredat room temperature for 12 hours. The reaction mixture was diluted withEtOAc (50 mL), filtered and concentrated. The residue was dissolved intoEtOAc (100 mL) and washed with water (100 mL) and brine (50 mL) anddried (MgSO₄). After filtration and concentration, the residue waspurified by chromatography on silica gel (0 to 5% EtOAc/Hexanes) to givethe title compound 4 (532 mg, 74%). [M+H]⁺: 267.0

(E)-4-(4-(1-methyl-1H-pyrazol-4-yl)but-3-en-1-ynyl)benzoic acid (IC-11)

Reagent MW Eq. g/ml mmol ACHL059-4 266.29 1.0 1.00 g 3.74 2M NaOH aq.40.00 2.0  2.4 mL 4.8 THF   25 mL MeOH   25 mL

The product (650 mg, 71%) was made and separated using the GeneralMethod for basic hydrolysis. [M+1]⁺: 369.0. ¹H-NMR: (400 MHz, DMSO-d₆):δ 3.83 (s, 3H), 6.29 (d, J=16.0 Hz, 1H), 7.00 (s, J=16.0 Hz, 1H), 7.55(d, J=8.4 Hz, 2H), 7.75 (s, 1H), 7.92-7.96 (m, 3H).

Synthesis of 4-(4-methylpent-3-en-1-ynyl)benzoic acid (IC-12)

Methyl 4-(4-methylpent-3-en-1-ynyl)benzoate (2)

To a solution of methyl 4-ethynylbenzoate (3.84 g, 24 mmol), Pd(PPh₃)₄(2.22 g, 1.92 mmol), CuI (0.32 g, 1.68 mmol) and PPh₃ (1.13 g, 4.32mmol) in degassed piperidine (160 mL) was added1-bromo-2-methylprop-1-ene (6.38 g, 47.26 mmol) at room temperature.After the mixture was heated at 60° C. for 3 h, the reaction wasquenched by sat. NH₄Cl solution and then was extracted with ethylacetate twice. The combined organic layer was washed with brine anddried over anhydrous sodium sulfate. After filtration of the dryingagent, the filtrate was evaporated, and the residue was purified bysilica-gel column (1% ethyl acetate in petroleum) to give compound 2(1.9 g, the yield was 37%) as a light yellow solid. ¹H NMR (400 MHz,CDCl₃): δ ppm, 1.88 (s, 3H), 1.99 (s, 3H), 3.91 (s, 3H), 5.49 (s, 1H),7.46 (d, J=8.4 Hz, 2H), 7.96 (d, J=8.4 Hz, 2H).

4-(4-methylpent-3-en-1-ynyl)benzoic acid (IC-12)

The compound (IC-12)(1.45 g, 82%) as a white solid was made andseparated using the General Method for basic hydrolysis. ¹H NMR (400MHz, DMSO-d₆): δ ppm, 1.87 (s, 3H), 1.96 (s, 3H), 5.59 (s, 1H), 7.52 (d,J=8.8 Hz, 2H), 7.91 (d, J=8 Hz, 2H), 13.01 (s, 1H)

The following compounds were prepared by using the correspondingintermediate acids synthesized as described above.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 108-1

329 108-2

343 108-3

383 108-4

381 108-5

327 108-6

329

Example 109 Synthesis of4-(2-fluoro-4-(4-(morpholinomethyl)phenyl)but-1-en-3-ynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)benzamide(109-1)

Synthesis of 4-(2-bromo-2-fluorovinyl)benzonitrile (1)

The THF solution (10 ml) of p-cyanobenzaldehyde (1.31 g, 10.0 mmol),triphenylphosphine (3.14 g, 12 mmol) and Tribromofluromethane (3.25 g,12 mmol) was cooled to 0° C. The THF solution (1 ml) of Diethyl zinc(1.47 g, 12 mmol) was added drop wise over 20 min. The reaction waswarmed to r.t and stirred overnight. The reaction mixture was cooled to0° C. and aqueous NH₄Cl (Sta'd, 20 ml) was added to quench the reaction.The mixture was extracted with Ethyl Acetate (100 ml×3). The combinedorganic layers was washed with HCl (1N, 100 ml), H₂O (100 ml×2) andbrine (100 ml). The crude product was purified with ISCO normal phasesilica gel column (0-10% EtOAc/Hex) to give desired 1.33 g product 1 asa trans/cis mixture (1:1) in the yield of 58.8%.

Synthesis of 4-(2-fluoro-4-(4(morpholinomethyl)phenyl)-but-1-en-3-ynyl)benzonitrile

The THF solution (5 ml) of compound 1 (226 mg, 1.0 mmol) and 2 (201 mg,1.0 mmol) was charged N₂ gas for 15 min. PdCl₂ (PPh₃)₂ (21 mg, 0.03mmol), CuI (19.0 mg, 0.1 mmol) and TEA (0.5 mml) were added to reactionsequentially at r.t. The reaction was stirred at r.t. overnight. Dilutedwith EtOAc, the reaction mixture was filtered through celite. Thefiltrate was washed with brine (50 ml) and concentrated. The crudeproduct was purified with ISCO normal phase silica gel column (0-50%EtOAc/Hex) to give 300 mg compound 3 in the yield of 86.7% as a 1:1mixture of trans and cis against F-double bond.

Synthesis of 4-(2-fluoro-4-(4(morpholinomethyl)phenyl)-but-1-en-3-ynyl)benzoic acid (4)

Compound 3 (300 mg, 0.87 mmol) was dissolved in 10 ml Dioxin and 5 mlH₂O. LiOH (104 mg, 4.3 mmol) was added to the solution. The reaction washeated to 120° C. under microwave for 2 hr. after cooling to r.t, thereaction mixture was acidified with 1N HCl to pH=3˜4. Diluted with H₂O,the mixture was extracted with EtOAc to give 240 mg compound 4 as crudeproduct in the yield of 75.5%.

Synthesis of 4-2-fluoro-4-(4-(morpholinomethyl)phenyl)but-1-en-3-ynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)benzamide(6)

DIPEA (0.67 ml, 3.85 mmol) was added to the DMF solution (5 ml) ofcompound 4 (240 mg, 0.77 mmol) and 5 (211 mg, 1.15 mmol) and HATU (380mg, 1.0 mmol) at r.t. the reaction was stirred at r.t overnight. H₂O wasadded to quench the reaction. The reaction mixture was extracted withEtOAc (50 ml×3). The combined organic layers was washed with H₂O (50ml×2) and brine (50 ml) then dried with Na₂SO₄. The crude product waspurified with ISCO normal phase silica gel column (0-10% MeOH/DCM) togive compound 6 (140 mg, 37.8%) as a 1:1 mixture of trans and cisagainst F-double bond

Synthesis of4-(2-fluoro-4-(4-(morpholinomethyl)phenyl)but-1-en-3-ynyl)-N-((2S,3R)-3-hydroxy-1-(hydroxyamino)-1-oxobutan-2-yl)benzamide(109-1)

Aqueous NH2OH (50% in H₂O, 0.5 ml) was added to the IPA solution (4 ml)of compound 6 (140 mg, 0.29 mmol) at r.t. The reaction was stirredovernight at r.t. After removing the reaction solvent, the crude productwas purified by prep. HPLC to give compound (109-1), a mixture of transand cis against double bond, (13 mg, 7.4%, m+z=482.2) as TFA salt.

Com- Scale Yield Yield Purity Retention pound (mmol)* (mg) (%)* (%) [M +H] time (min)** 109-1 0.29 13 mg 7.4% 94.4 492.2 1.508/1.634

Synthesis of (2S,3R)-methyl2-(4-((Z)-3-fluoro-4-(4-(morpholinomethyl)phenyl)but-3-en-1-ynyl)benzamido)-3-hydroxybutanoate109-2

Synthesis of 4-(2-bromo-2-fluorovinyl)benzaldehyde (2)

Compound 1 (417 mg, 1.85 mmol) was dissolved in DCM/Tol mixed solvent (6ml, 1:1). The solution was cooled to −78 C. DIBAL (2.2 ml, 2.21 mmol,1.0M in DCM) was added dropwise. The reaction was stirred at −78 C for 2hrs. NH4Cl (50 ml, aq. sta'd) was added to quench the reaction and themixture was stirred at r.t. for 2 hrs. The reaction mixture wasextracted with EtOAc (50 ml×3). The combined organic layers was washedwith H₂O (50 ml×2) and brine (50 ml). The crude product was purifiedwith ISCO normal phase silica gel column (0-20% EtOAc/Hex) to give 250mg compound 2 (250 mg, 59%) as 1:1trans/cis mixture against double bond.

Synthesis of (E)-4-(4-(2-bromo-2-fluorovinyl)benzyl)morpholine (3)

NaBH(OAc)₃ (693 mg, 3.27 mmol) was added to the THF solution (5 ml) ofcompound 2 (250 mg, 1.09 mmol) and morphline (190 mg, 2.18 mmol). Thereaction was stirred at r.t. overnight. THF was removed and the residuewas dissolved in 50 ml EtOAc and 50 ml H₂O. After separation, the waterlayer was extracted with EtOAc (50 ml×2). The combined organic layerswas washed with H₂O (50 ml×2) and brine (50 ml) and dried with Na₂SO4.the crude the product was purified with ISCO normal phase silica gelcolumn (0-50% EtOAc/DCM) to give compound 3 (209 mg, 63.9%) as a mixtureof trans/cis in the ration of 7:1 based on NMR.

Synthesis of methyl4-(3-fluoro-4-(4-(morpholinomethyl)phenyl)but-3-en-1-ynyl)benzoate (4 &5)

Compound 3 (210 mg, 0.67 mmol) and methyl 4-ethynylbenzoate (119 mg,0.75 mmol) were dissolved in 5 ml degassed THF. PdCl₂ (PPh₃)₂ (14 mg,0.02 mmol), CuI (13 mg, 0.07 mmol) and TEA (0.3 ml, 2.0 mmol) were addedsequentially to the reaction at r.t. The reaction was stirred at R.T.for 4 hrs. The reaction mixture was filtered. The filtered solid waswashed with EtOAc (100 ml). The organic solution was washed with H₂O (50ml×2) and brine (50 ml) and dried with Na₂SO₄. The crude product waspurified with ISCO normal phase silica gel column (0-50% EtOAc/DCM) togive compound 4 (197 mg) and 5 (33 mg) in 81.4% total yield.

Synthesis of(Z)-4-(3-fluoro-4-(4-(morpholinomethyl)phenyl)but-3-en-1-ynyl)benzoicacid (6)

Compound 4 (197 mg, 0.52 mmol) was dissolved in THF/MEOH (4 ml, 1:1).The aq. solution (2 ml) of LiOH (25 mg, 1.04 mmol) was added to thereaction. Reaction was stirred at r.t overnight. Diluted with H₂O (10ml), the reaction mixture was neutralized with HOAc to pH=7 andextracted with EtOAc (20 ml×5) to give 180 mg crude product 6 in theyield of 94.8%.

Synthesis of (2S,3R)-methyl2-(4-((Z)-3-fluoro-4-(4-(morpholinomethyl)phenyl)but-3-en-1-ynyl)benzamido)-3-hydroxybutanoate7

Reagent MW Eq. g/ml mmol Compound 6 365.4 1 180 mg 0.493 Threonine169.61 1.1 99.3 mg 0.54 HATU 380 1.2 225 mg 0.59 DIPEA 129 4.5 0.38 ml2.2 DMF 5 ml

The compound 7 in the yield of 86.6% was prepared using the GeneralMethod for HATU coupling.

Synthesis of (2S,3R)-methyl2-(4-((Z)-3-fluoro-4-(4-(morpholinomethyl)phenyl)but-3-en-1-ynyl)benzamido)-3-hydroxybutanoate109-2

Reagent MW Eq. g/ml Mmol Compound 7 480.53 1 221 mg 0.44 NH₂OH (50% inH₂O) 1.1 ml IPA 2 ml

The compound 109-2 (70 mg, 30.7%, m+z=482.2) as HCl salt was made usingthe General Method for hydroxamate formation.

The following compound was synthesized as described above.

Compound Amount # Structure obtained Yield M + Z 109-3

0.7 mg TFA salt 0.1% 482.2

Example 110 Synthesis of(E)-4-(4-(4-(morpholinomethyl)phenyl)but-1-en-3-ynyl)benzoic acid (1)

The detail procedure refers to Example 30 for synthesis of alternativeintermediate 009Each of the following compounds were prepared by the same syntheticroute as described in Example 96 and as for compound 110-12 as describedin Example 97

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 110-1(same as 30-12a)

464.5 3.62 A 110-2

462.4 5.19 B 110-3

463.4 6.11 B 110-4

504.4 4.67 B 110-5

464.4 3.61 A 110-6

478.5 4.79 B 110-7

478.5 4.79 B 110-8

552.5 4.50 B 110-9

460.3 4.84 B 110-10

501.1 4.23 B 110-11

496.4 5.57 B 110-12

488.4 5.41 B

Example 111N-[2-Amino-1-((S)-hydroxycarbamoyl)-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-1-en-3-ynyl]-benzamide(111-1)N-[2-(2-dimethylamino-acetylamino)-1-((S)-hydroxycarbamoyl)-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-1-en-3-ynyl]-benzamide(111-2)N-[2-acetylamino-1-((S)-hydroxycarbamoyl)-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-1-en-3-ynyl]-(111-3)N-[1-((S)-Hydroxycarbamoyl)-2-methylamino-ethyl]-4-[(E)-4-(4-morpholin-4-ylmethyl-phenyl)-but-1-en-3-ynyl]-benzamide(111-4)

The following compounds were prepared by the same synthetic route asdescribed in Examples 98 and 99.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 111-1

449.4 3.46 B 111-2

534.3 3.64 B 111-3

491.4 4.17 B 111-4

463.5 3.59 B

Example 112 Synthesis of (E)-methyl4-(4-(4-(hydroxymethyl)phenyl)but-1-en-3-ynyl)benzoate (1)

The detail procedure refers to Example 30 for synthesis of alternativeintermediate 011Each of the following compounds was synthesized using the syntheticroute as described in Example 101.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 112-1

394.5 4.59 B 112-2

380.0 4.53 B 112-3

395.2 5.29 B 112-4

409.4 5.99 B 112-5

409.4 5.57 B

Example 113

Each of the following compounds was synthesized using the syntheticroute as described in Example 102 and for compound 113-11 as describedin Example 103

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 113-1

419.4 3.86 B 113-2

433.4 3.79 B 113-3

434.5 4.49 B 113-4

434.4 4.56 B 113-5

449.0 4.98 A 113-6

447.1 3.95 B 113-7

448.3 5.01 B 113-8

511.3 3.88 B 113-9

408.3 6.22 B 113-10

422.4 6.65 B 113-11

477.5 4.74 B

Example 114 Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-((E)-4-phenyl-but-1-en-3-ynyl)-benzamide

Synthesis of 4-((E)-2-Iodo-vinyl)-benzoic acid methyl ester (2)

The title compound 2 (3.8 g) as a slightly yellow solid was prepared byusing the procedure of compound 4 of Example 96. Check of a sample byHPLC-MS showed the product peak (Ret. time (Method C) 3.10 min,MH⁺=289.2). This material could be further purified by silica gelchromatography using hexanes or pentane with 0-30% EtOAc as eluant.

Synthesis of 4-((E)-4-Phenyl-but-1-en-3-ynyl)-benzoic acid methyl ester(4)

The 4-((E)-2-iodo-vinyl)-benzoic acid methyl ester (403 mg, 1.40 mmol,1.0 eq), phenylacetylene* (157 mg, 1.54 mmol, 1.1 eq), copper(I) iodide(10.7 mg, 0.056 mmol, 0.04 eq) and PdCl₂(dppf) (22.9 mg, 0.028 mmol,0.02 eq) were dissolved with THF (4.5 mL) and diisopropylamine (396 uL,2.8 mmol, 2.0 eq) and sealed under nitrogen. The reaction mixture wasthen stirred rapidly for 1 min and placed in the microwave reactor at80-120° C. for 6-18 min until the vinyl iodide was consumed (HPLC-MS).The reaction mixture was then stirred and evaporated under a nitrogenstream to a slurry, and diluted with EtOAc (25 mL) and water (10 mL).After mixing and adjusting to pH˜5 with 3 M HCl, the aqueous phase wasremoved. The organic phase was washed with 0.3 M HCl (15 mL, 2×), andsatd aq NaCl (10 mL). Filtration of the brown solution through Na₂SO₄and evaporation of the solvents yielded the title compound as a crudeproduct (˜450 mg). This material could be further purified by silica gelchromatography using hexanes or pentane with 0-100% EtOAc as eluant, andevaporation of the title compound as a yellow glass. Check of a sampleby HPLC-MS showed a major product peak (Ret. time (Method C) 3.53 min,MH⁺=263.3). *—For aliphatic alkynes, using 3 eq of alkyne is required toobtain similar yields.

Synthesis of 4-((E)-4-Phenyl-but-1-en-3-ynyl)-benzoic acid (5)

Reagent MW Eq. mmol mg, ml 4-((E)-4-Phenyl-but-1-en-3-ynyl)- 262.31 1.0~1.4 ~367 mg benzoic acid methyl ester (4) THF 3.6 mL MeOH 1.2 mL 1M aqNaOH 2.0 2.8 2.8 mL

The 4-((E)-4-phenyl-but-1-en-3-ynyl)-benzoic acid methyl ester washydrolyzed and the acid precipitated using the “General Method for basichydrolysis”. Check of a sample by HPLC-MS showed a major product peak(Ret. time (Method A) 5.79 min, [MH+DMSO]⁺=327.1).

Synthesis of(2S,3R)-3-(9H-Fluoren-9-ylmethoxycarbonylamino)-2-[4-((E)-4-phenyl-but-1-en-3-ynyl)-benzoylamino]-butyricacid methyl ester (6)

Reagent MW, d Eq. mmol mg, ml 4-((E)-4-Phenyl-but-1-en-3- 248.28 1.00.30 74.5 mg ynyl)-benzoic acid (5) DIEA 129.24, 0.742 3.0 0.90 157 ulHATU 380.23 1.25 0.375 157 mg DMF 1.2 ml N-Fmoc-(S)-MeDAP-OMe × 390.871.12 0.336 131 mg HCl

The 4-((E)-4-phenyl-but-1-en-3-ynyl)-benzoic acid was coupled toFmoc-Me-DAP using the “General Method for HATU coupling”. Check of asample by HPLC-MS showed a major product peak (Ret. time (Method C) 3.57min, MH⁺=585.4).

Synthesis of (2S,3R)-3-Amino-2-[4-((E)-4-phenyl-but-1-en-3--benzoylamino]-butyric acid methyl ester (7)

The FMOC group of (2S,3R)-3-(9H-fluoren-9-ylmethoxycarbonylamino)-2-[4-((E)-4-phenyl-but-1-en-3-ynyl)-benzoylamino]-butyric acidmethyl ester was deprotected by dissolving in 50% piperidine in THF (6mL), stirring for 1-8 h, and evaporating the volatiles under vacuum or anitrogen stream to yield the desired product residue which may beoptionally purified by silica gel chromatography using hexanes with0-100% EtOAc as eluant. Check of a sample by HPLC-MS showed a majorproduct peak (Ret. time (Method C) 2.51 min, MH⁺=363.2).

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-((E)-4-phenyl-but-1-en-3-ynyl)-benzamide(114-1)

The (2S,3R)-3-amino-2- [4-((E)-4-phenyl-but-1-en-3-ynyl)-benzoyl amino]butyric acid methyl ester was dissolved in dioxane (0.6 mL), isopropanol(2.4 mL) and 50% aq hydroxylamine (3 mL), and optionally a catalyticamount of potassium cyanide (2 mg) was added at room temperature andstirred for 8-24 h until disappearance of the methyl ester (HPLC-MS).The reaction mixture was diluted in dichloromethane (8 mL), and theorganic layer separated, dried (Na₂SO₄), and evaporated on the rotovapto yield a slurry that was dissolved in DMSO. After subjecting thesample to preparative HPLC, a major peak was collected which showed anHPLC-MS of the desired product (Ret. time (Method A) 4.10 min,MH⁺=364.2). A second minor peak from preparative HPLC could also becollected as an isomer of the hydroxamic acid.

Each of the following compounds was synthesized as described above usingthe appropriate alkyne.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 114-1

363.4 6.2  B 114-2

382.3 6.59 B 114-3

382.3 6.23 B 114-4

397.9 7.13 B 114-5

397.9 6.73 B 114-6

432.3 7.54 B 114-7

447.9 7.71 B 114-8

365.1 2.38 B 114-9

365.1 2.67 B 114-10

398.4 4.56 AEach of the following compounds of was synthesized as described aboveusing the appropriate alkyne, threonine methyl ester hydrochloride or(S)-Me₂-BOC-DAP-OMe and BOC deprotection.

Ret. HPLC- Compound MH⁺ Time MS # Structure (m/z) (min) Method 114-11

383.1 7.21 B 114-12

383.1 7.01 B 114-13

366.3 3.30 B 114-14

365.3 7.01 B 114-15

328.7 3.27 A 114-16

378.8 6.48 B 114-17

370.3 6.99 B 114-18

342.3 5.54 B 114-19

344.3 6.09 B 114-20

370.3 6.88 B

General Procedure for Following Examples Method 6 (Sonogashira Coupling)

A microwave tube was charged with bromo- or iodo-phenyl derivative (1.9mmol), 4-ethynyl-benzoic acid methyl ester or acid (2.0 mmol),PdCl₂(dppf).CH₂Cl₂ (0.038 mmol, 2 mol %), CuI (0.076 mmol, 4 mol %),DIPA (4 mL), and THF (12 mL). The tube was backfilled with nitrogen,sealed, and irradiated in a microwave reactor (max. power 250 W) at100-125° C. for 10-20 min. THF was removed in vacuo and the residue wastaken up in EtOAc (100 mL). The organic layer was washed with water(2×50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo. Crude product was purified by normal phase flashchromatography on a CombiFlash® Companion™ unit using hexanes-EtOAcgradient elution. Fractions containing the desired product were combinedand concentrated to give the target compound (Fmoc protecting groups arecleaved during the reaction).

Example 115N-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-((4-((4-fluoropiperidin-1-yl)methyl)phenyl)ethynyl)benzamide(115-1)

Synthesis of 4-ethynyl-benzoic acid (1)

A mixture of 4-ethynylbenzonitrile (1.27 g, 10 mmol) and 2M aq. LiOH (25ml, 50 mmol) in dioxane (20 ml) was irradiated in microwave oven (max.power 250 W, 120° C.) for 30 min and cooled to ambient temperature.Reaction mixture was diluted with water (300 ml) and acidified with 1 Maq. HCl to pH˜3. Formed precipitate was filtrated and washed with waterand ether and dried in vacuum overnight to produce hydrochloric salt oftarget material (2.30 g, 63%) as yellow solid.

Synthesis of 4-(4-formyl-phenylethynyl)-benzoic acid (2)

A solution of compound 1 (1.46 g, 10 mmol), 4-iodobenzaldehyde (2.32 g,10 mmol) and diisopropylamine (3 ml) in THF (6 ml) was purged for 10 minwith dry nitrogen. A mixture of PdCl₂(PPh₃)₂ (210 mg, 0.03 mmol), andCuI (114 mg, 0.06 mmol) was added and reaction mixture was stirred atambient temperature overnight. Formed precipitate was filtrated.Filtrate was evaporated in vacuum. Residue was dissolved in EtOAc (100ml) and stirred with 5% aq. NaHCO₃ (30 ml). Formed precipitate wasfiltrated, washed with water, EtOAc, ether and dried in vacuum toprovide target product (2.4 g, 96%) as off-white solid.

Synthesis of 4-[4-(4-fluoro-piperidin-1-ylmethyl)-phenylethynyl]-benzoicacid (3)

A mixture of compound 2 (150 mg, 0.6 mmol), 4-fluoropiperidinehydrochloride (84 mg, 0.6 mmol), AcOH (30 μl) and silica-supportedNaCNBH₃ (120 mg, 0.12 mmol) in EtOH (1 ml) was irradiated in microwaveoven (max. power 250 W, 120° C.) for 25 min and cooled to ambienttemperature. Solids were filtrated. Filtrate was evaporated. Residue wasdissolved in EtOAc (100 ml), washed with 5% aq. NaHCO₃ (30 ml) and brine(30 ml), dried over anh. Na₂SO₄ and evaporated in vacuum. Residue wasdried in vacuum overnight to provide target material (68 mg, 34%) aswhite solid. LC-MS [M+H] 338.4.

Synthesis of (2S,3R)-3-azido-2-tert-butoxycarbonylamino-butyric acidmethyl ester (4)

A solution of di-tert-butyl dicarbonate (810 mg, 3.7 mmol),(2S,3R)-2-amino-3-azido-butyric acid methyl ester hydrochloride (660 mg,3.38 mmol) and DIEA (645 μl, 3.7 mmol) in i-PrOH (10 ml) was maintainedat ambient temperature for 10 h. Reaction mixture was evaporated invacuum. Residue was dissolved in water and extracted with hexane/ether(1:1). Combined organics were dried over anhydrous MgSO₄ andconcentrated in vacuum. Residue was subjected to flash chromatography onCombiFlash® Companion unit equipped with RediSep® flash column (normalphase, 35-60 micron average particle size silicagel, 40 g, TeledyneIsco); flow rate=35 ml/min; injection volume 2 ml; mobile phase A:hexane; mobile phase B: EtOAc; gradient 0-50% B in 1 h. Fractionscontaining the desired product were combined and concentrated in vacuumto provide target compound (564 mg, 65%) as colorless oil. LC-MS [M+H]259.0.

Synthesis of (2S,3R)-3-amino-2-tert-butoxycarbonylamino-butyric acidmethyl ester (5)

Compound 4 (786 mg, 3.05 mmol) was dissolved in methanol (20 ml)followed by the addition of Pd/C (5% wt, 200 mg). Reaction mixture wassubjected to hydrogenation (Parr apparatus, 80 psi) at ambienttemperature for 40 min. Catalyst was filtered and washed with methanol.Filtrate was evaporated in vacuum to provide 3 (682 mg, 96%) ascolorless oil. LC-MS [M+H] 233.0.

Synthesis of(2S,3R)-2-tert-butoxycarbonylamino-3-(9H-Fluoren-9-ylmethoxycarbonyl-amino)-butyricacid methyl ester (6)

A mixture of 5 (682 mg, 2.94 mmol) and Fmoc-OSu (1.04 g, 3.08 mmol) inacetone (5 ml) was stirred at ambient temperature for 2 h. Solvent wasevaporated in vacuum. Residue was dissolved in EtOAc (50 ml) and washedwith 5% NaHCO₃, and brine. Organic layer was dried over Na₂SO₄ andevaporated in vacuum. Residue was subjected to flash chromatography onCombiFlash® Companion unit equipped with RediSep® flash column (normalphase, 35-60 micron average particle size silicagel, 40 g, TeledyneIsco); flow rate=35 ml/min; injection volume 2 ml; mobile phase A:hexane; mobile phase B: EtOAc; gradient 0-45% B in 1 h. Fractionscontaining the desired product were combined and concentrated in vacuumto provide 6 (1.10 g, 82%) as colorless oil that was solidified duringvacuum drying. LC-MS [M+H] 455.3.

Synthesis of(2S,3R)-2-amino-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyric acidmethyl ester hydrochloride (7)

Compound 6 (1.10 g, 2.42 mmol) was dissolved in 4 N HCl/dioxane (8 ml)and solution was maintained at ambient temperature for 20 min. Theresulting suspension was diluted with ether and the precipitate wasfiltered and washed with ether. Compound was dried in vacuum to providethe hydrochloride of 7 (840 mg, 89%) as white solid. LC-MS [M+H] 355.2.

Synthesis of(2S,3R)-3-(9H-fluoren-9-ylmethoxycarbonylamino)-2-{4-[4-(4-fluoro-piperidin-1-ylmethyl)-phenylethynyl]-benzoylamino}-butyricacid methyl ester (8)

A solution of 3 (31 mg, 0.09 mmol), HATU (34 g, 0.09 mmol) and DIEA (50μl, 0.27 mmol) in DMF (1 ml) was maintained at ambient temperature for10 min followed by the addition of compound 7 hydrochloride (35 mg, 0.09mmol). Reaction mixture was stirred at ambient temperature overnightfollowed by the dilution with EtOAc (100 ml). Solution was extractedwith water (20 ml×2) and brine (20 ml). Organic layer was dried overMgSO₄ and evaporated. Residue was dried in vacuum to provide targetcompound (41 mg, 67%) as brown solid. LC-MS [M+H] 674.6.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(4-fluoro-piperidin-1-ylmethyl)-phenylethynyl]-benzamide(115-1)

A solution of hydroxylamine hydrochloride (26 mg, 0.36 mmol) in MeOH (2ml) was cooled to −5° C. followed by the addition of 25% NaOMe/MeOH (125μl, 0.55 mmol) under nitrogen. Reaction mixture was stirred at −5° C.for additional 5 min, cooled to −20° C., and solution of 8 (41 mg, 0.061mmol) in THF/MeOH (1:1, 2 ml) was added dropwise over the period of 5min. Temperature of reaction mixture was raised to ambient. Completionof the reaction was monitored by LC-MS. Reaction mixture was acidifiedwith 1 N aq. HCl to pH-′7 and evaporated in vacuum. Residue wasdissolved in DMSO (500 μl) and subjected to HPLC purification.[Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flow rate=20 ml/min;mobile phase A: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1%TFA; gradient elution from 8% B to 28% B in 60 min., detection 254 nm].Fractions containing the desired product were combined and lyophylisedto provide the trifluoracetate salt of 115-1 as white solid. LC-MS [M+H]452.9.

Retention Scale Yield Yield Purity time Compound (mmol) (mg) (%) (%)[M + H] (min)* 115-1 0.061 15.6 37 99.1 452.9 2.58 *[Chromolith SpeedRodRP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobile phase A:0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elution from 5% Bto 100% B over 9.6 min, detection 254 nm]The following compounds were made as described for 115-1 above.

Compound # Structure RT (min) [M + H] 115-2

5.11¹ 407.1 115-3

5.61¹ 471.5 115-4

2.59² 450.3 115-5

2.67² 472.3 115-6

3.01² 437.1 115-7

3.31² 514.3 115-8

3.54² 465.1 ¹Using LC-MS Analytical Method D. ²Using LC-MS AnalyticalMethod B.

Example 116(S)—N-(3-amino-1-(hydroxyamino)-1-oxopropan-2-yl)-4-((4-((2-aminoacetamido)methyl)phenyl)ethynyl)benzamide(116-1)

Synthesis of(S)-2-[4-(4-Aminomethyl-phenylethynyl)-benzoylamino]-3-(9H-fluoren-9-ylmethoxycarbonylamino)-propionicacid methyl ester hydrochloride (2)

Compound 1 (500 mg, 1.42 mmol), DAP (Fmoc, OMe).HCl (589 mg, 1.56 mmol),HATU (597 mg, 1.57 mmol), DIEA (0.791 mL, 4.54 mmol) and DMF (2 mL) werecombined and the mixture was stirred at ambient temperature for 30 min.Reaction mixture was diluted with EtOAc (100 mL), washed with water(2×50 mL) and brine (50 mL), dried over anhydrous MgSO₄, andconcentrated in vacuo. LC-MS: RT (Method A) 7.37 min; [M+H] 674.4. Theresidue was dissolved in dioxane (10 mL) and 4 M HCl/dioxane (25 mL, 100mmol) was added. Reaction mixture was stirred at ambient temperature for1 h, and volatiles were removed in vacuo to give target compound 2(1.134 g, 131%). LC-MS: RT (Method A) 5.07 min; [M+H] 574.5.

Synthesis of(S)-2-(4-{4-[(2-tert-Butoxycarbonylamino-acetylamino)-methyl]-phenylethynyl}-benzoylamino)-3-(9H-fluoren-9-ylmethoxycarbonylamino)-propionicacid methyl ester hydrochloride (3)

Compound 2 (365 mg, 0.598 mmol), Boc-Gly-OH (126 mg, 0.719 mmol), HATU(274 mg, 0.721 mmol), DIEA (0.365 mL, 2.09 mmol) and DMF (1 mL) werecombined and the mixture was stirred at ambient temperature for 1 h.Reaction mixture was diluted with EtOAc (100 mL), washed with water(2×50 mL) and brine (50 mL), dried over anhydrous MgSO₄, andconcentrated in vacuo. LC-MS: RT (Method A) 6.36 min; [M+H] 731.5(C₄₂H₄₂N₄O₈+H, requires 731.84). The residue was dissolved in dioxane (5mL) and 4 M HCl/dioxane (15 mL, 60 mmol) was added. Reaction mixture wasstirred at ambient temperature for 1 h, and volatiles were removed invacuo to give target compound 3 (489 mg, 122%). LC-MS: RT (Method A)5.23 min; [M+H] 631.5.

(S)—N-(3-amino-1-(hydroxyamino)-1-oxopropan-2-yl)-4-((4-((2-aminoacetamidomethyl)phenyl)ethynyl)benzamide (116-1)

Reagent MW Eq. mg/mL mmol Compound 3 667.17 1.0 147 mg 0.220Hydroxylamine hydrochloride 69.49 6 92 mg 1.32 25% NaOMe/MeOH 54.02 120.607 mL 2.66 THF 2 mL MeOH 2 mL

The target compound 116-1 (3.1 mg, 2.2% yield) as a white solid wasprepared by following General Method for hydroxamate. LC-MS: RT (MethodA) 2.39 min; [M+H] 410.5.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 116-1 0.220 3.1 2.2 99.1 410.4 2.39 ¹Based on the amountof compound used in the last step of the reaction. ²Using LC-MSAnalytical Method A.

Example 117N-((1S,2R)-2-Amino-1-methylcarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(117-1) and(2S,3R)-3-Amino-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-butyricacid (117-2)

Synthesis of(2S,3R)-3-(9H-Fluoren-9-ylmethoxycarbonylamino)-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-butyricacid methyl ester (2)

Compound 1 (137 mg, 0.383 mmol), MeDAP (Fmoc, OMe).HCl (150 mg, 0.384mmol), HATU (173 mg, 0.455 mmol), DIEA (0.298 mL, 1.71 mmol) and DMF (1mL) were combined and the mixture was stirred at ambient temperature for1 h. Reaction mixture was diluted with EtOAc (50 mL), washed with water(2×30 mL) and brine (30 mL), dried over anhydrous MgSO₄, andconcentrated in vacuo to give target compound 2 (314 mg, 125%). LC-MS:RT (Method A) 5.35 min; [M+H] 658.4.

Synthesis ofN-((1S,2R)-2-Amino-1-methylcarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(117-1)

A solution of compound 2 (˜0.19 mmol) in 2 M NH₂Me/THF (2 mL) wasstirred at 80° C. in a sealed reaction vessel for 16 h. Reaction mixturewas concentrated in vacuo and the residue was purified by preparativescale reverse-phase HPLC (Phenomenex Gemini C-18 column, 110 Å, 30×100mm, flow rate: 20 mL/min, mobile phase A: 0.1% TFA/water, mobile phaseB: 0.1% TFA/ACN, gradient elution from 5% B to 40% B over 90 min, MSdetection). Fractions containing the desired product were combined andlyophilized to provide the trifluoroacetate salt of target compound117-1 (10.8 mg, 8.58% yield) as a white solid. LC-MS: RT (Method A) 2.80min; [M+H] 435.4.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 117-1 0.19 10.8 8.58 98.5 435.4 2.80 ¹Based on the amountof compound used in the last step. ²Using LC-MS Analytical Method A.

Synthesis of(2S,3R)-3-Amino-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-butyricacid (117-2)

To a solution of compound 2 (˜0.19 mmol) in THF (1 mL), was added 2 MNaOH (aq) (0.475 mL, 0.950 mmol), and the mixture was stirred at ambienttemperature for 1 h. Reaction mixture was neutralized with 1 M HCl (aq)and concentrated in vacuo. The residue was purified by preparative scalereverse-phase HPLC (Phenomenex Gemini C-18 column, 110 Å, 30×100 mm,flow rate: 20 mL/min, mobile phase A: 0.1% TFA/water, mobile phase B:0.1% TFA/ACN, gradient elution from 5% B to 40% B over 60 min, MSdetection). Fractions containing the desired product were combined andlyophilized to provide the trifluoroacetate salt of target compound117-2 (26.7 mg, 21.6% yield) as a white solid. LC-MS: RT (Method A) 2.42min; [M+H] 422.1.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 117-2 0.19 26.7 21.6 98.9 422.1 2.42 ¹Based on the amountof compound used in the last step. ²Using LC-MS Analytical Method A.

Example 118N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-azetidin-1-ylmethyl-phenylethynyl)-benzamide(118-1)

Synthesis of(2S,3R)-3-(9H-Fluoren-9-ylmethoxycarbonylamino)-2-[4-(4-hydroxymethyl-phenylethynyl)-benzoylamino]-butyricacid methyl ester (2)

Compound 1 (615 mg, 2.44 mmol), MeDAP (Fmoc, OMe).HCl (954 mg, 2.44mmol), HATU (928 mg, 2.93 mmol), DIEA (1.4 mL, 8.04 mmol) and DMF (5 mL)were combined and the mixture was stirred at ambient temperature for 1h. Reaction mixture was diluted with EtOAc (100 mL), washed with water(2×50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give target compound 2 (1.429 g, 99.6%). LC-MS:RT (Method A) 6.19 min; [M+H] 589.3.

Synthesis of(2S,3R)-2-[4-(4-Azetidin-1-ylmethyl-phenylethynyl)-benzoylamino]-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (3)

To a stirred solution of compound 2 (1.429 g, 2.43 mmol) and DIEA (1.27mL, 7.29 mmol) in DCM (10 mL), cooled in ice/salt bath, was addeddropwise methanesulfonyl chloride (0.571 mL, 7.38 mmol). Reactionmixture was allowed to attain ambient temperature and stir for 30 min.One sixth of the reaction mixture (˜1.8 mL, ˜0.41 mmol) was addeddropwise to a solution of azetidine (0.111 mL, 1.64 mmol) and DIEA(0.286 mL, 1.64 mmol) in DCM (1 mL). Reaction mixture was stirred atambient temperature for 16 h, then diluted with EtOAc (100 mL), washedwith water (2×50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give target compound 3 (247 mg, 96%). LC-MS: RT(Method A) 2.90 min; [M+H] 628.4.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-azetidin-1-ylmethyl-phenylethynyl)-benzamide(118-1)

Reagent MW Eq. mg/mL mmol Compound 3 627.75 1.0 247 mg 0.393Hydroxylamine hydrochloride 69.49 6 164 mg 2.36 25% NaOMe/MeOH 54.02 121.08 mL 4.72 THF 2 mL MeOH 2 mL

The target compound 118-1 (3.0 mg, 1.2% yield) as a white solid wasprepared by following General Method for hydroxamate. LC-MS: RT (MethodA) 2.54 min; [M+H] 407.3.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 118-1 0.393 3.0 1.2 97.1 407.3 2.54 ¹Based on the amountof compound used in the last step of the reaction. ²Using LC-MSAnalytical Method A.Each of the following compounds was synthesized as described above.

Compound # Structure RT¹ (min) [M + H] 118-2

2.56 473.2 118-3

2.76 457.1 118-4

2.90 437.4 118-5

2.61 451.5 118-6

2.82 435.3 118-7

2.38 451.3 118-8

2.25 420.3 118-9

2.27 412.9  118-10

5.18 490.3 ¹Using LC-MS Analytical Method A.

Example 119N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-{4-[(2,2-difluoro-ethylamino)-methyl]-phenylethynyl}-benzamide(119-1)

Synthesis of(3S,4R)-3-(9H-Fluoren-9-ylmethoxycarbonylamino)-2-[4-(4-formyl-phenylethynyl)-benzoylamino]-butyricacid methyl ester (2)

Compound 1 (448 mg, 1.79 mmol; see Example A), MeDAP (Fmoc, OMe).HCl(700 mg, 1.79 mmol), HATU (816 mg, 2.15 mmol), DIEA (1.09 mL, 6.26 mmol)and DMF (3 mL) were combined and the mixture was stirred at ambienttemperature for 1 h. Reaction mixture was diluted with EtOAc (50 mL),washed with water (2×30 mL) and brine (30 mL), dried over anhydrousMgSO₄, and concentrated in vacuo to give target compound 2 (1.533 g,146%). LC-MS: RT (Method A) 7.06 min; [M+H] 587.2.

Synthesis of(2S,3R)-2-(4-{4-[(2,2-Difluoro-ethylamino)-methyl]-phenylethynyl}-benzoylamino)-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (3)

To a stirred solution of compound 2 (147 mg, 0.251 mmol) and2,2-difluoroethylamine (27 mg, 0.337 mmol) in THF (2 mL), was addedNaBH(OAc)₃ (74 mg, 0.349 mmol), and the mixture was stirred at ambienttemperature for 72 h. LC-MS analysis of the reaction mixture showed only10% conversion to desired product. 2,2-Difluoroethylamine (20 mg, 0.251mmol), acetic acid (0.1 mL, 1.75 mmol) and NaBH(OAc)₃ (100 mg, 0.472mmol) were added, and the reaction mixture was stirred at ambienttemperature for 2 h. LC-MS analysis of the reaction mixture showedcomplete conversion to desired product. Reaction mixture was quenchedwith 5% NaHCO₃ (aq) (50 mL) and extracted with EtOAc (3×20 mL) Combinedorganic layers were washed with water (30 mL) and brine (30 mL), driedover anhydrous MgSO₄, and concentrated in vacuo to give target compound3 (178 mg, 109%). LC-MS: RT (Method A) 5.60 min; [M+H] 652.3.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-{4-[(2,2-difluoro-ethylamino)-methyl]-phenylethynyl}-benzamide(119-1)

Reagent MW Eq. mg/mL mmol Compound 3 627.75 1.0 — ~0.251 Hydroxylaminehydrochloride 69.49 6 105 mg 1.51 25% NaOMe/MeOH 54.02 12 0.685 mL 3.00THF 2 mL MeOH 2 mL

The target compound 119-1 (20.1 mg, 12.2% yield) as a white solid wasprepared by following General Method for hydroxamate. LC-MS: RT (MethodA) 2.38 min; [M+H] 431.1 (C₂₂H₂₄F₂N₄O₃+H, requires 431.47).

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 119-1 0.251 20.1 12.2 99.2 431.1 2.38 ¹Based on theamount of compound used in the reductive amination. ²Using LC-MSAnalytical Method A.The following compounds were made as described above.

Compound # Structure RT (min) [M + H] 119-2

2.81¹ 421.2 119-3

2.35¹ 461.2 119-4

3.51² 432.3 119-5

3.78² 446.3 119-6

2.91² 395.1 ¹Using LC-MS Analytical Method A. ²Using LC-MS AnalyticalMethod B.

Example 120N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-ethoxymethyl-phenylethynyl)-benzamide(120-1)

Synthesis of 1-Ethoxymethyl-4-ethynyl-benzene (2)

To a solution of 4-ethynylbenzyl alcohol (436 mg, 3.30 mmol) in THF (2mL) was added NaH (101 mg, 4.21 mmol) and iodoethane (0.486 mL, 6.08mmol). Reaction mixture was stirred at ambient temperature for 16 h.Reaction mixture was diluted with brine (50 mL) and extracted with EtOAc(2×50 mL). Combined organic layers were washed with brine (50 mL), driedover anhydrous MgSO₄, and concentrated in vacuo. Residue was purified bynormal phase flash chromatography on a CombiFlash® Companion™ unitequipped with Luknova flash column (80 g silica gel, 40-60 μM averageparticle size, 60 Å pore size); flow rate: 60 mL/min; mobile phase A:hexane; mobile phase B: EtOAc; gradient elution from 0% B to 30% B over60 min. Fractions containing the desired product were combined andconcentrated in vacuo to give target compound 2 (290 mg, 55%). LC-MS: RT(Method A) 5.16 min; compound not significantly ionizable.

Synthesis of 4-(4-Ethoxymethyl-phenylethynyl)-benzoic acid (3)

Compound 2 (271 mg, 1.69 mmol), 4-iodobenzoic acid (419 mg, 1.69 mmol),PdCl₂(PPh₃)₂ (36 mg, 0.051 mmol), CuI (19 mg, 0.100 mmol), DIPA (4 mL,28.3 mmol), and THF (12 mL) were combined in a microwave tube. The tubewas backfilled with nitrogen, sealed, and irradiated in a microwavereactor (max. power 250 W) at 120° C. for 10 min. Reaction mixture wasdiluted with EtOAc (200 mL), washed with 2% H₂SO₄ (75 mL) and water (75mL), dried over MgSO₄ and concentrated in vacuo to give target compound3 (470 mg, 99%). LC-MS: RT (Method A) 5.79 min; compound notsignificantly ionizable.

Synthesis of (2S,3R)-2-[4-(4-Ethoxymethyl-phenyleth1)-benzoylamino]-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyric acidmethyl ester (4)

Compound 3 (115 mg, 0.410 mmol), MeDAP (Fmoc, OMe).HCl (161 mg, 0.412mmol), HATU (187 mg, 0.492 mmol), DIEA (0.235 mL, 1.34 mmol) and DMF(0.8 mL) were combined and the mixture was stirred at ambienttemperature for 1 h. Reaction mixture was diluted with EtOAc (50 mL),washed with water (2×30 mL) and brine (30 mL), dried over anhydrousMgSO₄, and concentrated in vacuo to give target compound 4 (230 mg,91%). LC-MS: RT (Method A) 7.45 min; [M+H] 617.2.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-ethoxymethyl-phenylethynyl)-benzamide(120-1)

Reagent MW Eq. mg/mL mmol Compound 4 616.72 1.0 230 mg 0.372Hydroxylamine hydrochloride 69.49 6 157 mg 2.26 25% NaOMe/MeOH 54.02 121.023 mL 4.48 THF 2 mL MeOH 2 mL

The target compound 120-1 (28.7 mg, 15.1% yield) as a white solid wasprepared by following General Method for hydroxamate. LC-MS: RT (MethodA) 4.06 min; [M+H] 396.2 (C₂₂H₂₅N₃O₄+H, requires 396.47).

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 120-1 0.372 28.7 15.1 99.7 396.2 4.06 ¹Based on theamount of compound used in the last step. ²Using LC-MS Analytical MethodA.The following compounds were synthesized according to procedures forcompound 5.

Compound # Structure RT (min) [M + H] 120-2

2.95¹ 430.1 120-3

4.22² 382.3 ¹Using LC-MS Analytical Method A. ²Using LC-MS AnalyticalMethod B.

Example 121N—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-(4-cyclopropylaminomethyl-phenylethynyl)-benzamide(121-1)

Synthesis of2-Benzyloxycarbonylamino-3-tert-butoxycarbonylamino-3-methyl-butyricacid (2)

Compound 1 (1.467, 3.86 mmol) was dissolved in EtOH (10 mL), 3 M NaOH(aq) (10 mL, 30 mmol) was added, and the mixture was heated at 70° C.for 1 h. Reaction mixture was concentrated in vacuo and residue wasdissolved in water (50 mL), cooled in ice/water bath, and acidified topH 2 with 10% H₃PO₄ (aq). Mixture was extracted with EtOAc (30 mL, 2×20mL). Combined organic layers were washed with water (30 mL) and brine(30 mL), dried over anhydrous Na₂SO₄, and concentrated in vacuo to givetarget compound 2 (0.977 g, 69.1%) as a viscous clear oil. LC-MS: RT(Method A) 4.96 min; [M+H] 367.3 (C₁₈H₂₆N₂O₆+H, requires 367.43). Crudeproduct was used in next synthetic step without additional purification.

Synthesis of2-Benzyloxycarbonylamino-3-tert-butoxycarbonylamino-3-methyl-butyri cacid pentafluorophenyl ester (3)

To a solution of compound 2 (967 mg, 2.64 mmol) and PFP (680 mg, 3.69mmol) in DCM (15 mL) was added DCC (653 mg, 3.16 mmol). Reaction mixturewas allowed to stir at ambient temperature for 3 h. Reaction mixture wasfiltered, and solids (DCU) were washed with DCM (3×5 mL). Combinedfiltrate and washes were concentrated in vacuo, and the residue waspurified by normal phase flash chromatography on a CombiFlash®Companion™ unit equipped with Luknova flash column (40 g silica gel,40-60 μm average particle size, 60 Å pore size); flow rate: 40 mL/min;mobile phase A: hexane; mobile phase B: EtOAc; gradient elution from 0%B to 40% B over 60 min. Fractions containing the desired product werecombined and concentrated in vacuo to give target compound 3 (1.025 g,72.9%) as a viscous clear oil. LC-MS: RT (Method A) 7.25 min; [M+H]533.2.

Synthesis of[3-((R)-4-Benzyl-2-oxo-oxazolidin-3-yl)-2-benzyloxycarbonylamino-1,1-dimethyl-3-oxo-propyl]-carbamicacid tert-butyl ester (4)

Compound 3 (1.025 g, 1.92 mmol), (R)-(+)-4-benzyl-2-oxazolidinone(0.5117 g, 2.89 mmol), LiBr (0.3344 g, 3.85 mmol), TEA (0.5366 mL, 3.85mmol), and THF (10 mL) were combined and allowed to stir at ambienttemperature for 14 h. Reaction mixture was concentrated in vacuo, andresidue was partitioned between water (50 mL) and EtOAc (30 mL). Aqueouslayer was extracted with EtOAc (2×20 mL). Combined organic layers werewashed with water (30 mL) and brine (30 mL), dried over anhydrousNa₂SO₄, and concentrated in vacuo to give target compound 4 (1.153 g,115%) as an off-white solid foam. LC-MS analysis (Method A) showeddiastereomers, compounds 5 and 6, with retention times RT₅=6.60 min,RT₆=6.78 min. Both with [M+H] 526.3 (C₂₈H₃₅N₃O₇+H, requires 526.62).

Separation of diastereomers,[(R)-1-((R)-4-Benzyl-2-oxo-oxazolidine-3-carbonyl)-2-tert-butoxycarbonylamino-2-methyl-propyl]-carbamicacid benzyl ester (5) and[(S)-1-((R)-4-Benzyl-2-oxo-oxazolidine-3-carbonyl)-2-tert-butoxycarbonylamino-2-methyl-propyl]carbamicacid benzyl ester (6)

Method 1:

Crude compound 4 (1.153 g) was purified and its component diastereomerswere separated by normal phase flash column chromatography on aCombiFlash® Companion™ unit equipped with Luknova flash column (80 gsilica gel, 40-60 μm average particle size, 60 Å pore size); flow rate:50 mL/min; mobile phase A: hexane; mobile phase B: EtOAc; gradientelution from 0% B to 10% B over 5 min, then 10% B to 25% B over 90 min.25 mL fractions. Fractions containing target compound withdiastereomeric excess (d.e.) greater than 95% were combined andconcentrated in vacuo to give products as white solid foams (789.9 mgtotal yield, 78.1% from 3). F104-110: compound 6, 106.3 mg, 96% d.e.F111-127: overlap, mixture of 5 and 6, 580.9 mg. F128-140: compound 5,102.7 mg, −96% d.e.

Method 2:

The residue from combined fractions 111-127 (580.9 mg), overlap frompurification by Method A, was subjected to preparative scalereverse-phase HPLC

(Varian Microsorb 100-10 C-18 column (50×300 mm), flow rate: 50 mL/min,mobile phase A: 0.1% TFA/water, mobile phase B: 0.1% TFA/ACN, gradientelution from 20% B to 95% B over 90 min, UV 254 nm monitor, 50 mLfractions). Fractions containing target compound with d.e. greater than95% were combined, neutralized with NaHCO₃ (aq), concentrated in vacuoto approximately one half of original volume, and extracted with EtOAc(3×1:2 vol. EtOAc:vol. aq). The combined organic layers were washed withwater (1:2 vol. H₂O:vol. organics) and brine (1:2 vol. brine:vol.organics), dried over anhydrous Na₂SO₄, and concentrated in vacuo togive products as white solid foams (441 mg total yield, 75.9% recovery).F56-58: compound 5, 153 mg, 99.7% d.e. F59+60: overlap, mixture of 5 and6, 181 mg. F61-63: compound 6, 107 mg, 97.7% d.e.

Synthesis of[(S)-2-Amino-3-((R)-4-benzyl-2-oxo-oxazolidin-3-yl)-1,1-dimethyl-3-oxo-propyl]-carbamicacid tert-butyl ester hydrochloride (7)

To a solution of compound 6 (207 mg, 0.394 mmol) (combined portions fromseparation methods 1 and 2, 96.7% d.e.) in MeOH (2.5 mL), were added 1 NHCl (0.394 mL) and 10% Pd/C (22.3 mg). Reaction vessel was evacuated andbackfilled with H₂ three times, and reaction mixture was allowed to stirunder an atmosphere of H₂ at ambient temperature for 1 h. Reactionmixture was filtered and concentrated in vacuo to give target compound 7(174.6 mg, 103%) as a viscous clear oil which crystallized uponstanding. LC-MS: RT (Method A) 4.11 min; [M+H] 392.4.

Synthesis of [(S)-3-((R)-4-Benzyl-2-oxo-oxazolidin-3-yl)-2-(4-{4-[(tert-butoxycarbonyl-cyclopropyl-amino)-methyl]-phenylethynyl}-benzoylamino)-1,1-dimethyl-3-oxo-propyl]-carbamicacid tert-butyl ester (9)

To a solution of compound 8 (167.7 mg, 0.428 mmol) and DIEA (0.355 mL,2.04 mmol) in DMF (5 mL) was added HATU (162.9 mg, 0.428 mmol), and themixture was stirred at ambient temperature for 5 min. Compound 7 (174.6mg, 0.408 mmol) was added, and the mixture was stirred at ambienttemperature for 20 min. Reaction mixture was added to 0.1 M HCl (aq) (80mL), and extracted with EtOAc (3×25 mL). Combined organic layers werewashed with water (40 mL) and brine (40 mL), dried over anhydrousNa₂SO₄, and concentrated in vacuo to give target compound 9 (351 mg,112%) as a sticky amber solid foam. LC-MS: RT (Method A) 8.60 min; [M+H]765.7.

Synthesis of [(S)-2-(4-{4-[(tert-Butoxycarbonyl-cyclopropyl-amino)-methyl]-phenylethynyl}-benzoylamino)-2-hydroxycarbamoyl-1,1-dimethyl-ethyl]-carbamicacid tert-butyl ester (10)

Compound 9 (0.408 mmol) was dissolved in dioxane (8 mL) with sonication,and 50% NH₂OH (aq) (2 mL) was added. Reaction mixture was allowed tostir at ambient temperature for 64 h. Reaction mixture was reduced toapproximately ⅓ of original volume by concentration in vacuo, dilutedwith water (50 mL), neutralized with 1 M HCl (aq), and extracted withEtOAc (3×30 mL). Combined organic layers were washed with water (40 mL)and brine (40 mL), dried over anhydrous Na₂SO₄, and concentrated invacuo to give target compound 10 as an amber solid foam. LC-MS: RT(Method A) 6.72 min; [M+H] 621.6.

Synthesis ofN—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-(4-cyclopropylaminomethyl-phenylethynyl)-benzamide(121-1)

To compound 10 was added 4 M HCl/dioxane (5 mL), and the mixture wasstirred at ambient temperature for 1 h. Volatiles were removed in vacuoto give a sticky amber solid residue. Crude product was purified bypreparative scale reverse-phase HPLC (Phenomenex Gemini C-18 column, 110Å, 30×100 mm, flow rate: 20 mL/min, mobile phase A: 0.1% TFA/water,mobile phase B: 0.1% TFA/ACN, gradient elution from 5% B to 20% B over90 min, MS detection). Fractions containing the desired product werecombined and lyophilized to provide the trifluoroacetate salt of targetcompound 121-1 (44.6 mg, 20.1% yield from 7) as a white solid. LC-MS: RT(Method A) 2.61 min; [M+H] 421.3.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 121-1 0.408 44.6 20.1 99.7 421.3 2.61 ¹Based on theamount of carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.The following compound was synthesized as described above for 121-1.

Compound Structure RT (min)¹ [M + H] 121-2

2.61 421.2 ¹Using LC-MS Analytical Method A.

Example 122 N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-{4-[(2-methoxy-ethylamino)-methyl]-phenylethynyl}-benzamide (122-1)

Synthesis of 4-{4-[(2-Methoxy-ethylamino)-methyl]-phenylethynyl}-benzoicacid methyl ester (2)

To a stirred suspension of compound 1 (200 mg, 0.751 mmol) and DIEA(0.654 mL, 3.76 mmol) in DCM (3 mL), cooled in ice/water bath, was addeddropwise methanesulfonyl chloride (0.070 mL, 0.901 mmol). After 5 minthe ice/water bath was removed and reaction mixture was stirred atambient temperature for 5 min. LC-MS: RT (Method A) 6.03 min; compoundnot significantly ionizable. Reaction mixture was added dropwise to asolution of 2-methoxyethylamine (0.5 mL, 5.83 mmol) in DMF (5 mL), andthe mixture was stirred at ambient temperature for 15 min. Reactionmixture was concentrated in vacuo, and residue was partitioned betweenwater (100 mL) and EtOAc (50 mL). Layers were separated and aqueouslayer was extracted with EtOAc (2×30 mL). Combined organic layers werewashed with water (50 mL) and brine (50 mL), dried over anhydrousNa₂SO₄, and concentrated in vacuo to give target compound 2 (257 mg,106%) as a viscous amber oil. LC-MS: RT (Method A) 4.19 min; [M+H]324.3. Crude product was used in next synthetic step without additionalpurification.

Synthesis of 4-(4-{[tert-Butoxycarbonyl-(2-methoxy-ethyl)-amino]-methyl}-phenylethynyl)-benzoicacid (3)

To compound 2 (0.751 mmol) dissolved in CHCl₃ (15 mL) were added DIEA(0.262 mL, 1.50 mmol) and Boc₂O (197 mg, 0.903 mmol), and the mixturewas stirred at ambient temperature for 20 min.* LC-MS analysis of thereaction mixture showed complete reaction. LC-MS: RT (Method A) 7.48min; [M+H] 424.4. Reaction mixture was concentrated in vacuo, andresidue was dissolved in EtOH (5 mL). NaOH (3M, aq) (2 mL, 6 mmol) wasadded, and the mixture was stirred at ambient temperature for 30 min.Reaction mixture was diluted with water (75 mL), cooled in ice/waterbath, acidified to pH 2 with 10% H₃PO₄ (aq), and extracted with EtOAc(40 mL, 2×20 mL). Combined organic layers were washed with water (40 mL)and brine (40 mL), dried over anhydrous Na₂SO₄, and concentrated invacuo to give target compound 3 (326 mg, 106% from 1) as an off-whitesolid. LC-MS: RT (Method A) 6.20 min; [M+H] 410.2. Crude product wasused in next synthetic step without additional purification. *Bocprotection was omitted for secondary amines.

Synthesis of(2S,3R)-3-tert-Butoxycarbonylamino-2-[4-(4-{[tert-butoxycarbonyl-(2-methoxy-ethyl)-amino]-methyl}-phenylethynyl)-benzoylamino]-butyricacid methyl ester (4)

To a solution of compound 3 (100 mg, 0.244 mmol) and DIEA (0.128 mL,0.735 mmol) in DMF (2 mL) was added HATU (111.4 mg, 0.293 mmol), and themixture was stirred at ambient temperature for 5 min. Methyl DAP.HCl(72.2 mg, 0.269 mmol) was added, and the mixture was stirred at ambienttemperature for 20 min. Reaction mixture was added to 0.1 M HCl (aq) (80mL), and extracted with EtOAc (3×25 mL). Combined organic layers werewashed with water (40 mL) and brine (40 mL), dried over anhydrousNa₂SO₄, and concentrated in vacuo to give target compound 4 (207.8 mg,136%) as a sticky amber solid. LC-MS: RT (Method A) 7.00 min; [M+H]624.4 (C₃₄H₄₅N₃O₈+H, requires 624.76). Crude product was used in nextsynthetic step without additional purification.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-{4-[(2-methoxy-ethylamino)-methyl]-phenylethynyl}-benzamide(122-1)

Reagent MW Eq. mg/mL mmol Compound 4 623.75 1.0 — 0.244 HCl, 4.0Msolution in 1,4-dioxane 36.46 82 5 mL 20 Isopropyl alcohol (IPA) 2 mLHydroxylamine, 50 wt. % solution in 33.03 133 2 mL 32.6 water

The target compound 122-1 (11.0 mg, 8.37% yield from 3) as a white solidwas prepared by following General Method for hydroxamate. LC-MS: RT(Method A) 2.35 min; [M+H] 425.2.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 122-1 0.244 11.0 8.37 96.7 425.2 2.35 ¹Based on theamount of carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.The following compounds were synthesized as described for 122-1.

Compound Structure RT (min) [M + H] 122-2

2.96¹ 451.5 122-3

2.41¹ 445.1 122-4

2.58¹ 499.2 122-5

3.21² 465.6 122-6

2.94² 427.6 122-7 (same as 121-1)

3.20² 421.1 122-8

3.77² 466.3 ¹Using LC-MS Analytical Method A. ²Using LC-MS AnalyticalMethod B.

Example 123N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-cyclopropoxymethyl-phenylethynyl)-benzamide(123-1)

Synthesis of 4-(4-Vinyloxymethyl-phenylethynyl)-benzoic acid methylester (1)

Pd(TFA)₂ (3.1 mg, 0.5 mol %) and bathophenanthroline (3.1 mg, 0.5 mol %)were dissolved in BVE (5 mL, 38.8 mmol) in a vial, and the mixture wasstirred at ambient temperature for 5 min. Compound 1 (500 mg, 1.88 mmol)and DIEA (0.0164 mL, 0.094 mmol) were added, vial was backfilled withnitrogen, capped, and heated at 75° C. for 20 h. Reaction mixture wasconcentrated in vacuo, and the residue was purified by normal phaseflash chromatography on a CombiFlash® Companion™ unit equipped withLuknova flash column (40 g silica gel, 40-60 μm average particle size,60 Å pore size); flow rate: 40 mL/min; mobile phase A: hexane; mobilephase B: EtOAc; gradient elution from 0% B to 40% B over 60 min.Fractions containing the desired product were combined and concentratedin vacuo to give target compound 2 (0.394 g, 71.8%) as an off-whitesolid. LC-MS: RT (Method A) 7.28 min; compound not significantlyionizable.

Synthesis of 4-(4-Cyclopropoxymethyl-phenylethynyl)-benzoic acid methylester (3)

To a stirred solution of compound 2 (150 mg, 0.513 mmol) and CH₂I₂(0.100 mL, 1.24 mmol) in anhydrous DCE (3 mL), under a nitrogenatmosphere, cooled in an ice/water bath, was cautiously addeddiethylzinc (1.0 M in hexanes; 1.13 mL, 1.13 mmol). After 5 min theice/water bath was removed and reaction mixture was allowed to stir atambient temperature for 20 h. Reaction mixture was added cautiously tosaturated NH₄Cl (aq) (50 mL) and extracted with DCM (3×20 mL). Combinedorganic layers were washed with 1 N HCl (30 mL), water (30 mL),saturated NaHCO₃ (aq) (30 mL) and brine (20 mL), dried over anhydrousNa₂SO₄, and concentrated in vacuo to give target compound 3 (180 mg,115%) as an off-white solid. LC-MS: RT (Method A) 7.30 min; compound notsignificantly ionizable. ¹H NMR (250 MHz, CDCl₃) δ 0.5-0.65 (4H, m), 3.3(1H, m), 3.9 (3H, s), 4.55 (2H, s), 7.3 (2H, d), 7.5-7.6 (4H, m), 8.0(2H, d).

Synthesis of 4-(4-Cyclopropoxymethyl-phenylethynyl)-benzoic acid (4)

To compound 3 (˜0.513 mmol) were added EtOH (5 mL), DCM (2 mL), and 3 MNaOH (aq) (2 mL, 6 mmol), and the mixture was stirred at ambienttemperature for 30 min. Reaction mixture was diluted with water (150mL), cooled in ice/water bath, acidified to pH 3 with 10% H₃PO₄ (aq),and extracted with EtOAc (3×30 mL). Combined organic layers were washedwith water (30 mL) and brine (30 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give to give target compound 4 (113.9 mg, 76%from 2) as a slightly off-white solid. LC-MS: RT (Method A) 6.06 min;compound not significantly ionizable.

Synthesis of(2S,3R)-3-tert-Butoxycarbonylamino-2-[4-(4-cyclopropoxymethyl-phenylethynyl)-benzoylamino]-butyricacid methyl ester (5)

To a solution of compound 4 (111 mg, 0.382 mmol) and DIEA (0.200 mL,1.15 mmol) in DMF (5 mL) was added HATU (174 mg, 0.458 mmol), and themixture was stirred at ambient temperature for 5 min. Methyl DAP.HCl(113 mg, 0.420 mmol) was added, and the mixture was stirred at ambienttemperature for 20 min. Reaction mixture was added to water (100 mL),and extracted with EtOAc (3×30 mL). Combined organic layers were washedwith water (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give target compound 5 (239.2 mg, 123%) as asticky amber solid foam. LC-MS: RT (Method A) 6.87 min; [M+H] 507.0(C₂₉H₃₄N₂O₆+H, requires 507.61). Crude product was used in nextsynthetic step without additional purification.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-cyclopropoxymethyl-phenylethynyl)-benzamide(123-1)

Reagent MW Eq. mg/mL mmol Compound 5 506.60 1.0 — ~0.382 HCl, 4.0Msolution in 1,4-dioxane 36.46 31.4 3 mL 12 Isopropyl alcohol (IPA) 4 mLHydroxylamine, 50 wt. % solution in 33.03 171 4 mL 65.3 water

The target compound 123-1 (67.5 mg, 33.9% yield from 4) as a white solidwas prepared by following General Method for hydroxamate. LC-MS: RT(Method A) 4.19 min; [M+H] 408.3.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 123-1 0.382 67.5 33.9 97.7 408.3 4.19 ¹Based on theamount of carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.

Example 124N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(2,2,2-trifluoro-ethoxymethyl)-phenylethynyl]-benzamide(124-1)

Synthesis of 4-(4-Methanesulfonyloxymethyl-phenyleth n 1)-benzoic acidmethyl ester (2)

To a suspension of compound 1 (200 mg, 0.751 mmol) and DIEA (0.262 mL,1.5 mmol) in DCM (5 mL), cooled in ice/water bath, was added dropwisemethanesulfonyl chloride (0.070 mL, 0.901 mmol) and the mixture wasstirred in ice/water bath for 5 min. Reaction mixture was allowed toattain ambient temperature and stir for 10 min. Reaction mixture wasdiluted with DCM (40 mL), and washed with water (2×30 mL). Combinedaqueous layers were back-extracted with DCM (2×20 mL). Combined organiclayers were dried over anhydrous Na₂SO₄, and concentrated in vacuo togive target compound 2 (254.5 mg, 98.4%) as an off-white solid. LC-MS:RT (Method A) 6.05 min; compound not significantly ionizable.

Synthesis of 4-[4-(2,2,2-Trifluoro-ethoxymethyl)-phenylethynyl]-benzoicacid (3)

Sodium (25 mg, 1.09 mmol) was dissolved completely in2,2,2-trifluoroethanol (8 mL, 111 mmol). Compound 2 (254.5 mg, 0.739mmol) was added, and the mixture was heated to reflux for 5 min. NaOH(3M, aq) (6 mL, 18 mmol) was added, and the mixture was heated at 70° C.for 1 h. Reaction mixture was diluted with water (150 mL), cooled inice/water bath, acidified to pH 3 with 10% H₃PO₄ (aq), and filtered.Solids were washed with water (3×50 mL) and dried by lyophilization togive target compound 3 (238.9 mg, 96.7%) as a slightly off-white solid.LC-MS: RT (Method A) 6.01 min; compound not significantly ionizable.

Synthesis of(2S,3R)-3-tert-Butoxycarbonylamino-2-{4-[4-(2,2,2-trifluoro-ethoxymethyl)-phenylethynyl]-benzoylamino}-butyricacid methyl ester (4)

To a solution of compound 3 (100 mg, 0.299 mmol) and DIEA (0.156 mL,0.897 mmol) in DMF (2 mL) was added HATU (136 mg, 0.359 mmol), and themixture was stirred at ambient temperature for 5 min. Methyl DAP.HCl(88.4 mg, 0.329 mmol) was added, and the mixture was stirred at ambienttemperature for 20 min. Reaction mixture was added to 0.1 M HCl (aq) (80mL), and extracted with EtOAc (3×25 mL). Combined organic layers werewashed with water (40 mL) and brine (40 mL), dried over anhydrousNa₂SO₄, and concentrated in vacuo to give target compound 4 (192 mg,117%) as a sticky light amber solid. LC-MS: RT (Method A) 6.81 min;[M+H] 549.5. Crude product was used in next synthetic step withoutadditional purification.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(2,2,2-trifluoro-ethoxymethyl)-phenylethynyl]-benzamide(124-1)

Reagent MW Eq. mg/mL mmol Compound 4 548.56 1.0 — 0.299 HCl, 4.0Msolution in 1,4-dioxane 36.46 53.5 4 mL 16 Isopropyl alcohol (IPA) 2 mLHydroxylamine, 50 wt. % solution in 33.03 109 2 mL 32.6 water

The target compound 124-1 (38.1 mg, 22.6% yield from 3) as a white solidwas prepared by following General Method for hydroxamate. LC-MS: RT(Method A) 4.35 min; [M+H] 449.9.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 124-1 0.299 38.1 22.6 95.8 449.9 4.35 ¹Based on theamount of carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.The following compounds were synthesized according to procedures abovefor 124-1.

Compound Structure RT (min)¹ [M + H] 124-2

4.25 410.1 124-3

3.98 432.3 ¹Using LC-MS Analytical Method A.

Example 125N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-cyclopropoxy-phenylethynyl)-benzamide(125-1)

Synthesis of 1-Bromo-4-(2-chloro-ethoxy)-benzene (1)

To a mixture of 4-bromophenol (2.0 g, 11.6 mmol) and K₂CO₃ (3.20 g, 23.2mmol) in DMF (50 mL) was added 3-chloroethyl p-toluenesulfonate (2.52mL, 13.9 mmol), and the mixture was stirred at 60° C. for 16 h. Reactionmixture was concentrated in vacuo, and the residue was partitionedbetween water (200 mL) and DCM (50 mL). Aqueous layer was furtherextracted with DCM (3×20 mL). Combined organic layers were washed withwater (30 mL) and brine (30 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give target compound 1 (3.12 g, 115%) as alight amber oil. LC-MS: RT (Method A) 5.83 min; compound notsignificantly ionizable. ¹H NMR (250 MHz, CDCl₃) δ 3.8 (2H, t), 4.2 (2H,t), 6.8 (2H, d), 7.4 (2H, d). Crude product was used in next syntheticstep without additional purification.

Synthesis of 1-Bromo-4-vinyloxy-benzene (2)

To a stirred solution of compound 1 (˜11.6 mmol) in THF (30 mL), cooledin an ice/water bath, was added potassium tert-butoxide (1.95 g, 17.3mmol) in three portions. After 10 min the ice/water bath was removed andreaction mixture was stirred at ambient temperature for 16 h. Reactionmixture was concentrated in vacuo and partitioned between water (150 mL)and DCM (50 mL). Aqueous layer was further extracted with DCM (3×30 mL).Combined organic layers were washed with water (40 mL) and brine (40mL), dried over anhydrous Na₂SO₄, and concentrated in vacuo. Residue waspurified by normal phase flash chromatography on a CombiFlash®Companion™ unit equipped with Luknova flash column (40 g silica gel,40-60 μm average particle size, 60 Å pore size); flow rate: 40 mL/min;mobile phase: hexane. Fractions containing the desired product werecombined and concentrated in vacuo to give target compound 2 (1.702 g,74% from 4-bromophenol) as a clear oil. LC-MS: RT (Method A) 5.95 min;compound not significantly ionizable. ¹H NMR (250 MHz, CDCl₃) δ 4.45(1H, dd), 4.75 (1H, dd), 6.6 (1H, dd), 6.85 (2H, d), 7.4 (2H, d).

Synthesis of 1-Bromo-4-cyclopropoxy-benzene (3)

To a stirred solution of compound 2 (1.702 g, 8.55 mmol) and CH₂I₂ (5.52mL, 68.4 mmol) in anhydrous DCE (75 mL), under a nitrogen atmosphere,cooled in an ice/water bath, was cautiously added diethylzinc (1.0 M inhexanes; 68.4 mL, 68.4 mmol). After 30 min the ice/water bath wasremoved and reaction mixture was allowed to stir at ambient temperaturefor 64 h. Reaction mixture was added cautiously to saturated NH₄Cl (aq)(200 mL), mixture was shaken and layers separated, and aqueous layer wasextracted with DCM (3×30 mL). Combined organic layers were washed with 1N HCl (100 mL), water (100 mL), saturated NaHCO₃ (aq) (100 mL) and brine(80 mL), dried over anhydrous Na₂SO₄, and concentrated in vacuo to givea light amber oil. Crude product was purified by normal phase flashchromatography on a CombiFlash® Companion™ unit equipped with Luknovaflash column (40 g silica gel, 40-60 μm average particle size, 60 Å poresize); flow rate: 40 mL/min; mobile phase: hexane. Fractions containingthe desired product were combined and concentrated in vacuo to givetarget compound 3 (1.310 mg, 71.9%) as a clear oil. LC-MS: RT (Method A)5.87 min; compound not significantly ionizable. ¹H NMR (250 MHz, CDCl₃)δ 0.75 (4H, m), 3.7 (1H, m), 6.9 (2H, d), 7.4 (2H, d).

Synthesis of 4-(4-Cyclopropoxy-phenylethynyl)-benzoic acid methyl ester(4)

A microwave tube was charged with compound 3 (400 mg, 1.88 mmol),4-ethynyl-benzoic acid methyl ester (330 mg, 2.06 mmol),PdCl₂(dppf).CH₂Cl₂ (30.7 mg, 2 mol %), CuI (14.3 mg, 4 mol %), DIPA(0.40 mL, 2.83 mmol), and DMF (6 mL). Tube was backfilled with nitrogen,sealed, and irradiated in a microwave reactor (max. power 250 W) at 160°C. for 5 min. Reaction mixture was diluted with water (100 mL) andextracted with EtOAc (3×30 mL). Combined organic layers were washed withwater (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give a sticky brown solid. Crude product waspurified by normal phase flash chromatography on a CombiFlash®Companion™ unit equipped with Luknova flash column (40 g silica gel,40-60 μm average particle size, 60 Å pore size); flow rate: 40 mL/min;mobile phase A: hexane; mobile phase B: EtOAc; gradient elution from 0%B to 40% B over 60 min. Fractions containing the desired product werecombined and concentrated in vacuo to give target compound 4 (47.7 mg,8.7%) as a light brown solid. LC-MS: RT (Method A) 7.32 min; compoundnot significantly ionizable. ¹H NMR (250 MHz, CDCl₃) δ 0.75 (4H, m), 3.7(1H, m), 3.85 (3H, s), 7.0 (2H, d), 7.5 (2H, d), 7.6 (2H, d), 8.0 (2H,d).

Synthesis of 4-(4-Cyclopropoxy-phenylethynyl)-benzoic acid (5)

To compound 4 (47.7 mg, 0.163 mmol) were added EtOH (5 mL) and 3 M NaOH(aq) (2 mL, 6 mmol). Reaction mixture was heated at 70° C. for 30 min.Reaction mixture was diluted with water (150 mL), cooled in ice/waterbath, acidified to pH 3 with 10% H₃PO₄ (aq), and extracted with EtOAc(3×30 mL). Combined organic layers were washed with water (30 mL) andbrine (30 mL), dried over anhydrous Na₂SO₄, and concentrated in vacuo togive to give target compound 5 (44.7 mg, 98.8%) as a slightly off-whitesolid. LC-MS: RT (Method A) 6.11 min; compound not significantlyionizable.

Synthesis of(2S,3R)-3-tert-Butoxycarbonylamino-2-[4-(4-cyclopropoxy-phenylethynyl)-benzoylamino]-butyricacid methyl ester (6)

To a solution of compound 5 (44.7 mg, 0.161 mmol) and DIEA (0.084 mL,0.482 mmol) in DMF (2 mL) was added HATU (73.5 mg, 0.193 mmol), and themixture was stirred at ambient temperature for 5 min. Methyl DAP.HCl(47.6 mg, 0.177 mmol) was added, and the mixture was stirred at ambienttemperature for 20 min. Reaction mixture was added to water (100 mL),and extracted with DCM (3×30 mL). Combined organic layers were washedwith water (30 mL), dried over anhydrous Na₂SO₄, and concentrated invacuo to give target compound 6 (159.7 mg, 201%) as a viscous amber oil.LC-MS: RT (Method A) 6.89 min; [M+H] 493.2. Crude product was used innext synthetic step without additional purification.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-cyclopropoxy-phenylethynyl)-benzamide(125-1)

To compound 6 (˜0.161 mmol) was added 4 M HCl/dioxane (3 mL), and themixture was stirred at ambient temperature for 1 h. Volatiles wereremoved in vacuo, and the residue was washed with diethyl ether (2×10mL), centrifuging and removing supernatant after each wash, to give alight amber powdery solid. LC-MS: RT (Method A) 4.91 min; [M+H] 393.2(C₂₃H₂₄N₂O₄+H, requires 393.47). Isopropyl alcohol (4 mL) was added tothe solid and the mixture was cooled in an ice/water bath for 5 min.NH₂OH (50%, aq) (4 mL) was added to the mixture, dropwise for the first2 mL. Reaction mixture was allowed to stir in ice bath for 5 min, andthen allowed to stir at ambient temperature for 16 h. Solvent volume wasreduced approximately by half under a stream of nitrogen, and water (10mL) was added. The suspension was thoroughly agitated (vibro mixer andsonication), centrifuged and the supernatant was discarded. Water (10mL) was added to the solid and the suspension was thoroughly agitated,centrifuged and the supernatant was discarded. Wet solid was dried bylyophilization to give crude product (41.7 mg, 69.3% crude yield from 5)as an off-white solid. Crude product was purified by preparative scalereverse-phase HPLC (Phenomenex Gemini C-18 column, 110 Å, 30×100 mm,flow rate: 20 mL/min, mobile phase A: 0.1% TFA/water, mobile phase B:0.1% TFA/ACN, gradient elution from 15% B to 40% B over 60 min, MSdetection). Fractions containing the desired product were combined andlyophilized to provide the trifluoroacetate salt of target compound125-1 (30.2 mg, 37.0% yield from 5, 56.1% recovery from crude product)as a white solid. LC-MS: RT (Method A) 4.39 min; [M+H] 394.3.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 125-1 0.161 30.2 37.0 100 394.3 4.39 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.

Example 126N—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-(4-ethoxymethyl-phenylethynyl)-benzamide(126-1)

Synthesis of 4-(4-Ethoxymethyl-phenylethynyl)-benzoic acid (2)

To a solution of compound 1 (1.5 g, 5.63 mmol) in dry DMSO (30 mL) wasadded sodium hydride (2.25 g, 56.3 mmol), and the mixture was stirred atambient temperature for 10 min. Iodoethane (0.591 mL, 7.32 mmol) wasadded, and the mixture was stirred at ambient temperature for 1 h.Reaction mixture was diluted with water (150 mL), cooled in ice/waterbath, acidified to pH 3 with 1 M HCl (aq), and extracted with EtOAc (80mL, 2×50 mL). Combined organic layers were washed with water (80 mL) andbrine (80 mL), dried over anhydrous Na₂SO₄, and concentrated in vacuo togive a yellow solid. Crude product was purified by normal phase flashchromatography on CombiFlash® Companion™ unit equipped with Luknovaflash column (80 g silica gel, 40-60 μm average particle size, 60 Å poresize); flow rate: 50 mL/min; mobile phase A: CH₂Cl₂; mobile phase B:MeOH; gradient elution from 0% B to 8% B over 60 min. Fractionscontaining the desired product were combined and concentrated in vacuoto give target compound 2 (995 mg, 63%) as an off-white solid. LC-MS: RT(Method A) 5.89 min; compound not significantly ionizable. ¹H NMR (250MHz, DMSO-d6) δ 1.1 (3H, t), 3.6 (2H, q), 4.5 (2H, s), 7.4-8.0 (8H, m).

Synthesis of (S)-3-tert-Butoxycarbonylamino-2-[4-(4-ethoxymethyl-phenylethynyl)-benzoylamino]-3-methyl-butyric acid methylester (3)

To a solution of compound 2 (150 mg, 0.535 mmol) and DIEA (0.280 mL,1.61 mmol) in DMF (5 mL) was added HATU (244 mg, 0.642 mmol), and themixture was stirred at ambient temperature for 5 min. Chiral dimethylDAP (145 mg, 0.589 mmol) was added, and the mixture was stirred atambient temperature for 20 min. Reaction mixture was added to 0.1 M HCl(aq) (100 mL), and extracted with EtOAc (3×30 mL). Combined organiclayers were washed with water (50 mL) and brine (50 mL), dried overanhydrous Na₂SO₄, and concentrated in vacuo to give target compound 3(386 mg, 142%) as a viscous amber oil. LC-MS: RT (Method A) 7.20 min;[M+H] 509.3. Crude product was used in next synthetic step withoutadditional purification.

Synthesis ofN—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-(4-ethoxymethyl-phenylethynyl)-benzamide(126-1)

To compound 3 was added 4 B: 0.1% TFA/ACN, gradient elution from 20% Bto 55% B over 90 min, MS detection M HCl/dioxane (4 mL), and the mixturewas stirred at ambient temperature for 1 h. Volatiles were removed invacuo to give a sticky amber solid. LC-MS: RT (Method A) 4.70 min; [M+H]408.9 (C₂₄H₂₈N₂O₄+H, requires 409.51). Isopropyl alcohol (4 mL) wasadded to the solid and the mixture was cooled in an ice/water bath for 5min. NH₂OH (50%, aq) (4 mL) was added to the mixture, dropwise for thefirst 2 mL. Reaction mixture was allowed to stir in ice bath for 5 min,and then allowed to stir at ambient temperature for 16 h. Solvent volumewas reduced approximately by half under a stream of nitrogen, and water(10 mL) was added. The suspension was thoroughly agitated (vibro mixerand sonication), centrifuged and the supernatant was discarded. Water(10 mL) was added to the solid and the suspension was thoroughlyagitated, centrifuged and the supernatant was discarded. Wet solid wasdried by lyophilization to give crude product (160.7 mg, 73.3% crudeyield from 2) as an off-white solid. Crude product was purified bypreparative scale reverse-phase HPLC (Phenomenex Gemini C-18 column, 110Å, 30×100 mm, flow rate: 20 mL/min, mobile phase A: 0.1% TFA/water,mobile phase). Fractions containing the desired product were combinedand lyophilized to provide the trifluoroacetate salt of target compound126-1 (44.7 mg, 16% yield from 2, 22% recovery from crude product) as awhite solid. LC-MS: RT (Method A) 4.07 min; [M+H] 410.5 (C₂₃H₂₇N₃O₄+H,requires 410.50).

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 126-1 0.535 44.7 16 99.5 410.5 4.07 ¹Based on the amountof carboxylic acid used in the coupling reaction. ²Using LC-MSAnalytical Method A.Each of the following compounds was synthesized as described above forcompound 4.

Compound # Structure RT (min)¹ [M + H] 126-2

4.24 410.3 ¹Using LC-MS Analytical Method A.

Example 127N-[2-(Formyl-hydroxyamino)-ethyl]-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(127-1)

Synthesis N-(2-N,O-Di-Boc-hydroxyaminoethyl)phthalimide (1)

To the mixture of N-(2-bromoethyl)-phthalimide (473 mg, 1.86 mmol) andN,O-di-Boc-hydroxylamine (500 mg, 2.14 mmol) in DMF (5 ml) was addedK₂CO₃ (483 mg, 3.49 mmol). Reaction mixture was stirred at 65° C.overnight followed by the dilution with EtOAc (50 ml). Solution wasextracted with water (15 ml×2) and brine (30 ml). Organic layer wasdried over Na₂SO₄ and evaporated in vacuum. Residue was subjected toflash chromatography on CombiFlash® Companion unit equipped withRediSep® flash column (normal phase, 35-60 micron average particle sizesilicagel, 12 g, Teledyne Isco); flow rate=30 ml/min; injection volume 2ml; mobile phase A: hexane; mobile phase B: EtOAc; gradient 0-50% B in25 min. Fractions containing the desired product were combined andconcentrated in vacuum to provide target compound 450 mg (59.2%) ascolorless oil. LC-MS [M+H] 407.0.

Synthesis of 2-N,O-DiBoc-Hydroxyaminoethylamine (2)

A solution of compound 2 (450 mg, 1.11 mmol) and N₂H₄×H₂O (250 mg, 5.00mmol) in EtOH (5 ml) was heated at 80° C. for 1 h. The reaction mixturewas cooled to r.t., the formed white precipitant was filtrated andwashed with EtOH (5 ml). Filtrate was concentrated in vacuum and driedin vacuum for 4 h. Compound 2 was used as is for the next chemicaltransformation with no additional purification. LC-MS [M+H] 277.4.

Synthesis ofN-(2-N,O-di-Boc-hydroxyamino-ethyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(4)

A mixture of compound 2 (0.55 mmol), compound 3 (160 mg, 0.5 mmol), DIEA(330 μl, 2.0 mmol) and HATU (230 mg, 0.6 mmol) was stirred at ambienttemperature overnight followed by the dilution with EtOAc (50 ml) andextraction extracted with water (15 ml×2) and brine (30 ml). Organiclayer was dried over Na₂SO₄ and evaporated in vacuum. Residue wassubjected to flash chromatography on CombiFlash® Companion unit equippedwith RediSep® flash column (normal phase, 35-60 micron average particlesize silicagel, 4 g, Teledyne Isco); flow rate=18 ml/min; injectionvolume 2 ml; mobile phase A: hexane; mobile phase B: EtOAc; gradient0-30% B in 30 min. Fractions containing the desired product werecombined and concentrated in vacuum to provide target compound 250 mg(86.2%) as white solid. LC-MS [M+H] 480.2 (-Boc).

Synthesis ofN-(2-hydroxyamino-ethyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(5)

A solution of compound 4 (250 mg, 0.43 mmol) in of 4 M HCl/dioxane (5ml) was stirred at RT for 1 h. Solvents were evaporated in vacuum.Residue was dried in vacuum to provide dihydrochloride of compound 5(184 mg, 95%) as yellow solid. LC-MS [M+H] 380.1.

N-[2-(Formyl-hydroxyamino)-ethyl]-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(127-1)

A mixture of compound 5 (184 mg, 0.43 mmol), 2,2,2-triflouroethylformate (74 mg, 0.58 mmol), DIEA (165 μl, 1.0 mmol) in THF (5 ml) wasstirred at 65° C. for 2 h. The reaction mixture was concentrated invacuum, the residue was dissolved in DMSO (600 μl) and subjected to HPLCpurification. [Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flowrate=20 ml/min; mobile phase A: 100% water, 0.1% TFA; mobile phase B:100% ACN, 0.1% TFA; gradient elution from 10% B to 40% B in 60 min.,detection 254 nm]. Fractions containing the desired product werecombined and lyophilized to provide trifluoroacetic salt of targetcompound (40.3 mg, 17.9%) as white solid. LC-MS [M+H] 408.4.

Scale Yield Yield Purity Retention time Compound (mmol) (mg) (%) (%)[M + H] (min)* 127-1 0.43 40.3 17.9 95.7 408.4 3.15 *[ChromolithSpeedRod RP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobilephase A: 0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elutionfrom 5% B to 100% B over 9.5 min, detection 254 nm]

Example 128N-[2-(Formyl-hydroxy-amino)-1-hydroxymethyl-ethyl]-4-(4-morpholin-4-yl-methyl-phenylethynyl)-benzamide(128-1)

Synthesis of (2-tert-Butoxy-1-iodomethyl-ethyl)-carbamic acid benzylester (1)

A mixture of N-Cbz-O-tert-Bu-serinol (420 mg, 1.5 mmol), Ph₃P (487 mg,1.86 mmol) and imidazole (150 mg, 2.2 mmol) was dissolved in THF (10 ml)followed by the addition of I₂ (483 mg, 3.49 mmol) portion wise during 3min. The reaction mixture was stirred at ambient temperature overnightfollowed by the dilution with EtOAc (100 ml), and extraction with water(50 ml×3) and brine (30 ml). The organic layer was dried over Na₂SO₄ andevaporated in vacuum. The residue was used as is for the next chemicaltransformation with no additional purification. LC-MS [M+H⁺] 392.0.

Synthesis of(2-tert-Butoxy-1-(N,O-di-Boc-hydroxyaminomethyl-ethyl)-carbamic acidbenzyl ester (2)

A mixture of compound 1 (1.5 mmol), N,O-di-Boc-hydroxylamine (350 mg,1.5 mmol) and K₂CO₃ (483 mg, 3.49 mmol) in DMF (5 ml) was stirred at 65°C. for 2 h. The reaction mixture was diluted with EtOAc (50 ml), andextracted with water (15 ml×2) and brine (30 ml). The organic layer wasdried over Na₂SO₄ and solvent was removed in vacuum. Residue wassubjected to flash chromatography on CombiFlash® Companion unit equippedwith RediSep® flash column (normal phase, 35-60 micron average particlesize silicagel, 12 g, Teledyne Isco); flow rate=30 ml/min; injectionvolume 2 ml; mobile phase A: hexane; mobile phase B: EtOAc; gradient0-95% B in 60 min. Fractions containing the desired product werecombined and concentrated in vacuum to provide target compound (518 mg,73.3% overall for 2 steps) as colorless oil. LC-MS [M+H⁺] 497.2.

Synthesis of (2-tert-Butoxy-1-(N,O-di-Boc-hydroxyaminomethyl)-ethylamine(3)

A mixture of solution of compound 2 (518 mg, 1.04 mmol) in 15 ml of MeOHand 5% Pd/C (200 mg) was subjected to hydrogenation in Parr shaker at 65psi overnight. Pd/C was removed by filtration and filtrate wasconcentrated in vacuum to yield colorless oil, which was used on thenext step with no additional purification. LC-MS [M+H⁺] 363.4.

Synthesis ofN-(2-tert-Butoxy-1-(N,O-di-Boc-hydroxyaminomethyl)-ethyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(5)

A mixture of compound 3 (1.04 mmol), compound 4 (250 mg, 0.71 mmol),DIEA (496 μl, 3.0 mmol) and HATU (380 mg, 1.0 mmol) was stirred atambient temperature for 45 min followed by the dilution with EtOAc (50ml) and extraction with water (15 ml×2) and brine (30 ml). Organic layerwas dried over Na₂SO₄ and evaporated in vacuum. Residue was subjected toflash chromatography on CombiFlash® Companion unit equipped withRediSep® flash column (normal phase, 35-60 micron average particle sizesilicagel, 4 g, Teledyne Isco); flow rate=18 ml/min; injection volume 2ml; mobile phase A: hexane; mobile phase B: EtOAc; gradient 0-60% B in60 min. Fractions containing the desired product were combined andconcentrated in vacuum to provide target compound 5 250 mg (48.2%) aswhite solid. LC-MS [M+H] 666.1.

Synthesis ofN-(2-Hydroxyamino-1-hydroxymethyl-ethyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(6)

The mixture of compound 5 (0.37 mmol, 250 mg) and 4 N HCl/dioxane (6 ml,24 mmol) was stirred at r.t. for 1.5 h. Solvents were removed in vacuumand HCl salt was dissolved in a THF/DIEA mixture (10 ml/220 μl). A halfof resulting solution was used on next step. LC-MS [M+H⁺] 410.0.

N-[2-(Formyl-hydroxyamino)-ethyl]-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(128-1)

Reagent MW Eq. mg/ml mmol Compound 6 (as a solution, see pr. 276.34 1.0— 0.18 step.) Trifluoroethyl formate 128.05 11.1 195 mg 2.00 DIEA (as asolution, see pr. step.) 129.24 3.6 110 μl 0.65 THF (as a solution, seepr. step.) 5 ml

Trifluoroethyl formate (20 mg, 2 mmol) was added to the solution (˜5 ml)from previous step and the reaction mixture was stirred at 55° C. for 1h. The reaction mixture was concentrated in vacuum. Residue wasdissolved in DMSO (600 μl) and subjected to HPLC purification.[Phenomenex Gemini C-18 column, 110 Å (30×100 mm); flow rate=20 ml/min;mobile phase A: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1%TFA; gradient elution from 10% B to 40% B in 60 min., detection 254 nm].Fractions containing the desired product were combined and lyophilizedto provide trifluoroacetic salt of target compound (10.6 mg, 10.7%) aswhite solid.

Scale Yield Yield Purity Retention time Compound (mmol) (mg) (%) (%)[M + H] (min)* 128-1 0.18 10.6 10.7 97.7 438.1 2.94 *[ChromolithSpeedRod RP-18e C18 column (4.6 × 50 mm); flow rate 1.5 ml/min; mobilephase A: 0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient elutionfrom 5% B to 100% B over 9.5 mm, detection 254 nm]

Example 1293-Amino-N-hydroxy-3-methyl-2-[3-(4-morpholin-4ylmethyl-phenyl)-propynoylamino]-butyramide(129-1)

Synthesis of 3-Amino-2-[4-(4-hydroxymethyl-phenylethynyl)-benzoylamino]-3-methyl-butyric acidmethyl ester (2)

To a cold solution of compound 1 (700 mg, 1.7 mmol) in 6 mL NMP and 10mL AcOH of at 0° C., Zn dust (1.2 g, 18 mmol.) was added. The reactionmixture was vigorously stirred for 2 h at 25° C., monitored by LC-MSuntil there was no start material left. The reaction mixture wasfiltered though a celite pad, the solid was washed with DCM (3×10 mL)and MeOH (2×10 mL), combined the filtrate and washing liquid, and thenconcentrated to give 420 mg (65%) pure compound 2 as colorless oil.LC-MS: RT (Method A) 3.36 min; [M+H] 638.7.

Synthesis of3-tert-Butoxycarbonylamino-2-[4-(4-hydroxymethyl-phenylethynyl)-benzoylamino]-3-methyl-butyricacid methyl ester (3)

To a cold solution of compound 2 (320 mg, 0.84 mmol) in THF (4 mL) at 0°C., di-tert-butyl dicarbonate (200 mg, 0.92 mmol) was added. Thereaction mixture was stirred at ambient temperature for 2 h, until TLC(30% EtOAc in hexanes) indicated the disappearance of starting material.The reaction mixture was diluted with DCM (20 mL), washed with saturatedNaHCO₃, brine, dried over Na₂SO₄, and then concentrated to give thecrude compound 3 370 mg (92%).

Synthesis of3-tert-Butoxycarbonylamino-3-methyl-2-[4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzoylamino]-butyricacid methyl ester (5)

To a cold solution of compound 3 (370 mg, 0.77 mmol.) and DIEA (0.30 mL,1.6 mmol) was added MsCl (0.12 mL, 1.54 mmol.) at 0° C. The reactionmixture was stirred at ambient temperature for 30 min. until TLC (30%EtOAc in Hexanes) indicated the disappearance of starting material. Thereaction mixture was diluted with DCM (20 mL), washed with brine (2×20mL), dried over Na₂SO₄, concentrated to give the crude product 4 (400mg, 90%). The methanesulfonate 4 was re-dissolve in DCM (10 mL). To thesolution at 0° C., morpholine (0.35 mL, 4.0 mmol) was added dropwise.The mixture was stirred at ambient temperature for 30 min. Reactionmixture was concentrated in vacuo, and residue was partitioned betweenwater (30 mL) and EtOAc (30 mL). Layers were separated and aqueous layerwas extracted with EtOAc (2×10 mL). Combined organic layers were washedwith water (10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give compound 5 (439 mg, 100%) as a off-whitesolid.

Synthesis ofN-(2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide(6)

To a mixture of compound 5 (148 mg) in dioxane (4 mL), HCl (4 M indioxane, 4 mL) was added at 0° C. The reaction mixture was stirred atambient temperature for 1 h, until LC-MS indicated the disappearance ofstarting material. HCl and solvent were removed by vacuum; the residuewas dissolve in DCM, washed with sat. NaHCO₃, and brine, concentrated togive crude compound 6.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(2-cyclopropylamino-acetylamino)-phenylethynyl]-benzamide(129-1)

Compound 7 was made using the General Method for hydroxamate formation.Crude product was purified by preparative scale reverse-phase HPLC(Phenomenex Gemini C-18 column, 110 Å, 30×100 mm, flow rate: 20 mL/min,mobile phase A: 0.1% TFA/water, mobile phase B: 0.1% TFA/ACN, gradientelution from 15% B to 40% B over 60 min, MS detection). Fractionscontaining the desired product were combined and lyophilized to providethe trifluoroacetate salt of target compound 129-1 (11.4 mg, 6.2% yield)as a white solid. LC-MS: RT (Method A) 2.79 min; [M+H] 451.1.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 129-1 1 11.4 6.2 99.5 451.1 2.79 ¹Based on compound 5.²Using LC-MS Analytical Method A.

Example 130N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(2-cyclopropylamino-acetylamino)-phenylethynyl]-benzamide(130-1)

Synthesis of (4-Iodo-benzyl)-carbamic acid tert-butyl ester (1)

To a solution of 4-iodoaniline (1.95 g, 5.0 mmol) in iPrOH (10 mL) wasadded Boc₂O (1.199 g, 5.5 mmol), and the mixture was stirred at ambienttemperature for 16 h. iPrOH was removed in vacuo and the residue wascrystallized from Et₂O/hexanes mixture to give target compound 1 (1.219g, 76%) as a off-white solid. LC-MS: RT (Method A) 6.14 min; [2M+H]638.7.

Synthesis of 4-(4-tert-Butoxycarbonylamino-phenylethynyl)-benzoic acid(2)

Reagent MW Eq. mg, ml mmol 4-Ethynyl benzoic acid 146.15 1.0 146 mg 1.0Compound 1 319.14 1.0 319 mg 1.0 PdCl₂(PPh₃)₂ 703.91 0.03 21 mg 0.03 CuI190.45 0.06 12 mg 0.06 (iPr)₂NH 101.19 0.6 mL 0.3 THF 1.2 mL

The compound 2 was made using the General Method for Sonogashiracoupling, but without PPh₃. Yield: 355 mg. Crude product was used innext synthetic step without additional purification. LC-MS: RT (MethodA) 6.05 min, [M+H] 338.6.

Synthesis of(2S,3R)-2-[4-(4-tert-Butoxycarbonylamino-phenylethynyl)-benzoylamino]-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 2 337.38 1.0 355 mg 1.0 Fmoc-MethylDAP•HCl 390.87 1.0 390 mg 1.0 HATU 380.2 1.2 456 mg 1.2 DIEA 129.25 3.50.609 mL 3.5 DMF 1 mL

Compound 3 was made using the General Method for HATU coupling. LC-MS:RT (Method A) 7.49 min, [M+H] 674.2. Crude product was used in nextsynthetic step without additional purification.

Synthesis of(2S,3R)-2-[4-(4-Amino-phenylethynyl)-benzoylamino]-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (4)

Reagent MW Eq. mg/mL mmol Compound 3 673.77 1.0 — 1.0 HCl, 4.0M solutionin 1,4-dioxane 4 mL Dioxane 2 mL

Compound 4 was made using the General Method for Boc deprotection. Crudeproduct was used in next synthetic step without additional purification.

Synthesis of(2S,3R)-2-{4-[4-(2-Bromo-acetylamino)-phenylethynyl]-benzoylamino}-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (5)

To a suspension of compound 5 (137 mg, 0.22 mmol) and DIEA (96 uL, 0.55mmol) in CHCl₃ (1 mL), cooled in ice/water bath, was added dropwisebromoacetyl chloride (24 μL, 0.29 mmol) and the mixture was stirred inice/water bath for 5 min. Reaction mixture was allowed to attain ambienttemperature and stirred for 1.5 h. The reaction mixture was used on nextstep. LC-MS: RT (Method A) 6.91 min, MH⁺=695.5.

Synthesis of (2S,3R)-2-{4-[4-(2-Cyclopropylamino-acetylamino)-phenylethynyl]-benzoylamino}-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (6)

Cyclopropyl amine (24 μL, 0.35 mmol) was added to the reaction mixtureof compound 6 and resulting mixture was stirred for 1 h at ambienttemperature. The reaction mixture was diluted with EtOAc (80 mL) andorganic layer was washed with water (2×40 mL) and brine (40 mL), driedover anhydrous Na₂SO₄, and concentrated in vacuo to give target compound6 (133 mg, 90%) as a brown solid. LC-MS: RT (Method A) 6.82 min,MH⁺=671.3.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-(2-cyclopropylamino-acetylamino)-phenylethynyl]-benzamide(130-1)

Reagent MW, d Eq. mg, ml mmol Compound 6 670.77 1.0 133 mg 0.2 NH₂OH ×HCl 69.49 6.0 92 mg 0.78 25% MeONa in MeOH 12 0.6 mL 1.56 THF 2 mL MeOH2 mL

Compound 130-1 was made using the General Method for hydroxamateformation. LC-MS: RT (Method A) 2.48 min, MH⁺=450.2. Crude product waspurified by preparative scale reverse-phase HPLC (Phenomenex Gemini C-18column, 110 Å, 30×100 mm, flow rate: 20 mL/min, mobile phase A: 0.1%TFA/water, mobile phase B: 0.1% TFA/ACN, gradient elution from 15% B to40% B over 60 min, MS detection). Fractions containing the desiredproduct were combined and lyophilized to provide the trifluoroacetatesalt of target compound 130-1 (3.7 mg, 0.54% yield) as a white solid.LC-MS: RT (Method A) 3.08 min; [M+H] 449.9.

Scale Yield Yield Purity Retention Compound (mmol)¹ (mg) (%)¹ (%)² [M +H] time (min)² 130-1 1 3.7 0.54 95.0 449.9 3.08 ¹Based on the amount ofcarboxylic acid used in the coupling reaction. ²Using LC-MS AnalyticalMethod A.

Example 131N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-aminomethyl-phenylethynyl)-benzamide(131-1),N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-methanesulfonylamino-methyl)-phenylethynyl]-benzamide(131-2) andN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-guanidinomethyl-phenylethynyl)-benzamide(131-3)

Synthesis of (4-Iodo-benzyl)-carbamic acid tert-butyl ester (1)

To a solution of 4-iodobenzylamine (1.35 g, 5.0 mmol) and DIEA (0.96 mL,5.5 mmol) in DMF (5 mL) was added Boc₂O (1.09 g, 5.0 mmol), and themixture was stirred at ambient temperature for 1 h. The reaction mixturewas diluted with EtOAc (125 mL) and organic layer was washed with water(2×40 mL) and brine (40 mL), dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give target compound 1 (1.635 g, 98%) as aoff-white solid. LC-MS: RT (Method A) 6.02 min; [2M+H] 667.0. Crudeproduct was used in next synthetic step without additional purification.

Synthesis of4-[4-(tert-Butoxycarbonylamino-methyl)-phenylethynyl]-benzoic acidmethyl ester (2)

Reagent MW Eq. mg, ml mmol Methyl 4-Ethynyl benzoate 160.17 1.0 1635 mg4.9 Compound 1 333.17 1.1 864 mg 5.4 PdCl₂(PPh₃)₂ 703.91 0.03 104 mg0.15 PPh₃ 262.29 0.1 129 mg 0.49 CuI 190.45 0.06 56 mg 0.3 (iPr)₂NH101.19 3 mL THF 6 mL

The compound 2 was made using the General Method for Sonogashiracoupling. Crude product was used in next synthetic step withoutadditional purification. LC-MS: RT (Method A) 5.92 min, [M+H] 366.2).

Synthesis of4-[4-(tert-Butoxycarbonylamino-methyl)-phenylethynyl]-benzoic acid (3)

Reagent MW Eq. mg, ml mmol Compound 2 365.43 1.0 1.79 g 4.91 2N NaOH aq.160.17 6.0 15 mL 30 MeOH 100 mL THF 50 mL

All components were stirred for 16 h at room temperature. Most ofsolvent was removed in vacuo. Water (100 mL) was added to residue. Theaqueous solution was acidified with 2% H₂SO₄ aq. to pH=2-3 and extractedwith EtOAc (2×50 mL) The combined organic layer was washed with water(2×40 mL), brine (50 mL) and dried over Na₂SO₄. EtOAc was removed invacuo to give desired compound 3 (1.650 g, 96% based on compound 1) asan off-white solid. LC-MS: RT (Method A) 5.76 min; low ionization.

Synthesis of (2S,3R)-2-{4-[4-(tert-Butoxycarbonylamino-methyl)-phenylethynyl]-benzoylamino}-3(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (4)

Reagent MW Eq. mg/mL mmol Compound 3 351.41 1.0 235 mg 0.67 Fmoc-Methyl390.87 1.0 200 mg 0.67 DAP•HCl HATU 380.2 1.2 306 mg 0.8 DIEA 129.25 3.50.408 mL 2.35 DMF 1 mL

Compound 4 was made using the General Method for HATU coupling. LC-MS:RT (Method A) 7.54 min, [M+H] 688.2). Crude product was used in nextsynthetic step without additional purification.

Synthesis of(2S,3R)-2-[4-(4-Aminomethyl-phenylethynyl)-benzoylamino]-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid methyl ester (5)

Reagent MW Eq. mg/mL mmol Compound 4 687.8 1.0 — 0.67 HCl, 4.0M solutionin 1,4-dioxane 15 mL Dioxane  5 mL

Compound 5 was made using the General Method for Boc deprotection.LC-MS: RT (Method A) 5.47 min, [M+H] 588.3. Crude product was used innext synthetic step without additional purification.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-aminomethyl-phenylethynyl)-benzamide(131-1)

Reagent MW, d Eq. mg, ml mmol Compound 5 HCl 624.14 1.0 137 mg 0.22NH₂OH × HCl 69.49 6.0 92 mg 1.32 25% MeONa in MeOH 12 0.63 mL 2.64 THF 2mL MeOH 2 mL

Compound 131-1 was made using the General Method for hydroxamateformation. LC-MS: RT (Method A) 2.17 min, MH⁺=367.0.

Synthesis of(2S,3R)-3-(9H-Fluoren-9-ylmethoxycarbonylamino)-2-{4-[4-(methanesulfonylamino-methyl)-phenylethynyl]-benzoylamino}-butyricacid methyl ester (7)

To a suspension of compound 5 (137 mg, 0.22 mmol) and DIEA (77 μL, 0.44mmol) in CHCl₃ (0.5 mL), cooled in ice/water bath, was added dropwisemethanesulfonyl chloride (22 μL, 0.29 mmol) and the mixture was stirredin ice/water bath for 5 min. Reaction mixture was allowed to attainambient temperature and stir for 30 min. Reaction mixture was dilutedwith EtOAc (100 mL), washed with water (2×30 mL) and brine (25 mL).Combined organic layers were dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give target compound 7 (141 mg, 97%) as anoff-white solid. LC-MS: RT (Method A) 6.37 min, M+H⁺=666.4.

N-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-[4-methanesulfonyl-amino-methyl)-phenylethynyl]-benzamide(131-2)

Reagent MW, d Eq. mg, ml mmol Compound 7 665.77 1.0 141 mg 0.21 NH₂OH ×HCl 69.49 6.0 92 mg 1.32 25% MeONa in MeOH 12 0.63 mL 2.64 THF 2 mL MeOH2 mL

Compound 131-2 was made using the General Method for hydroxamateformation. LC-MS: RT (Method A) 3.13 min, MH⁺=445.3.

Synthesis of(2S,3R)-3-(9H-Fluoren-9-ylmethoxycarbonylamino)-2-[4-(4-guanidino-methyl-phenylethynyl)-benzoylamino]-butyricacid methyl ester (9)

To a suspension of compound 5 (137 mg, 0.22 mmol) and DIEA (191 μL, 1.1mmol) in DMF (0.5 mL), was added 1H-pyrazole-1-carboxamidine HCl (71 mg,0.48 mmol) and the mixture was stirred at 60° C. for 1 h. The reactionmixture was diluted with EtOAc (80 mL), washed with water (2×30 mL) andbrine (25 mL). Combined organic layers were dried over anhydrous Na₂SO₄,and concentrated in vacuo to give target compound 9 (82 mg, 65%) as anoff-white solid. LC-MS: RT (Method A) 5.48 min, MH⁺=630.3.

Synthesis ofN-((1S,2R)-2-Amino-1-hydroxycarbamoyl-propyl)-4-(4-guanidinomethyl-phenylethynyl)-benzamide(131-3)

Reagent MW, d Eq. mg, ml mmol Compound 9 629.72 1.0 82 mg 0.13 NH₂OH ×HCl 69.49 6.0 55 mg 0.78 25% MeONa in MeOH 12 0.35 mL 1.56 THF 2 mL MeOH2 mL

Compound 131-3 was made using the General Method for hydroxamateformation. LC-MS: RT (Method A) 2.72 min, MH⁺=409.2.

The following compounds were made as described above.

Ret. HPLC- Compound Time MH⁺ MS # Structure (min) (m/z) Method 131-1

2.58 367.1 A 131-2

4.23 445.5 A 131-3

2.99 409.1 A

Example 132N—[(S)-2-(2-Cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-phenylethynyl-benzamide(132-1)

Synthesis of(S)-3-tert-butoxycarbonylamino-2-(4-phenylethynyl-benzoylamino)-propionicacid methyl ester (2)

To the mixture of compound 1 (210 mg, 0.94 mmol) andH—(S)-DAP(N′-Boc)-OMe hydrochloride (275 mg, 1.08 mmol) in DMF (5 mL)was added HATU (429 mg, 1.13 mmol) followed by DIEA (491 μl, 2.82 mmol).Reaction mixture was stirred at ambient temperature for 20 min, dilutedwith 0.1 M HCl_(aq) (50 mL) and extracted with EtOAc (60 mL×2). Combinedorganic layers were washed with water (60 mL) and brine (60 mL), driedover anh. Na₂SO₄ and evaporated in vacuo to give target compound 2 (397mg, 100%) as an yellow oil. LC-MS [M+H] 424.0. Compound was used in nextsynthetic step without additional purification.

Synthesis of (S)-3-amino-2-(4-phenylethynyl-benzoylamino)-propionic acidmethyl ester (3)

To the solution of compound 2 (397 mg, 0.94 mmol) in dioxane (2 mL) andMeOH (2 mL) was added solution of 4 N HCl in dioxane (4 mL, 16 mmol).Reaction mixture was stirred at ambient temperature. Completion of thereaction was monitored by LC-MS. After completion (in 2 h) solvent wasevaporated in vacuo. Residue was dissolved in i-PrOH (10 mL) andevaporated in vacuo. The aforementioned procedure was repeated twice toprovide hydrochloride salt of target product 3 (337 mg, 100%) as a lightyellow solid. LC-MS [M+H] 323.7.

Synthesis of(S)-3-(2-bromo-acetylamino)-2-(4-phenylethynyl-benzoylamino)-propionicacid methyl ester (4)

To the mixture of compound 3 hydrochloride (337 mg, 0.94 mmol) and DIEA(410 μl, 2.35 mmol) in CH₂Cl₂ (8 mL) was added bromoacetyl bromide (82μl, 0.94 mmol) at 0° C. Reaction mixture was allowed to warm to ambienttemperature and stirred for 30 min. CH₂Cl₂ was evaporated in vacuo. Theresidue was dissolved in EtOAc (100 mL), washed with water (50 mL) andbrine (50 mL×2). Organic phase was dried over anh. Na₂SO₄ and evaporatedin vacuo. The crude product was purified by flash column chromatographyon silica gel utilizing a 5% MeOH/CH₂Cl₂ as an gradient to give thetarget product 4 (270 mg, 65%) as a white solid. LC-MS [M+H] 445.9.

Synthesis of(S)-3-(2-cyclopropylamino-acetylamino)-2-(4-phenylethynyl-benzoylamino)-propionicacid methyl ester (5)

To a solution of compound 4 (67 mg, 0.15 mmol) in CH₂Cl₂ (2 mL) wasadded cyclopropylamine (36 μl, 0.53 mmol) at 0° C. Reaction mixture wasmaintained at 0° C. for 10 min, mixture was then allowed to warm toambient temperature and stirred overnight. After completion the solventwas removed in vacuo. The residue was diluted with water (40 mL),acidified with 1 N HCl to pH 4 and extracted with EtOAc (40 mL×2).Combined organic layers were washed with water (50 mL) and brine (50mL), dried over anh. Na₂SO₄ and evaporated in vacuo to provide thetarget product 5 (63 mg, 100%) as a light yellow oil. LC-MS [M+H] 420.9.

Synthesis of N— [(S)-2-(2-cyclopropylamino-acetylamino)-1-hydroxycarbamoyl-ethyl]-4-phenylethynyl-benzamide(132-1)

To a stirred suspension of compound 5 (63 mg, 0.15 mmol) andhydroxylamine hydrochloride (83 mg, 1.2 mmol) in MeOH (anh, 2 mL) andTHF (anh, 0.8 mL) was added dropwise 25% NaOMe/MeOH (343 μl, 1.5 mmol)at −5° C. under nitrogen. Reaction mixture was stirred at −5° C. foradditional 5 min. Temperature of reaction mixture was raised to ambient.Completion of the reaction was monitored by LC-MS. After completion (in20 min) reaction mixture was acidified with 1 N HCl in MeOH to pH˜5 atlow temperature and evaporated in vacuo. Residue was dissolved in DMSO(400 μl) and subjected to HPLC purification. [Phenomenex Gemini C-18column, 110 Å (30×100 mm); flow rate=20 mL/min; mobile phase A: 100%water, 0.1% TFA; mobile phase B: 100% ACN, 0.1% TFA; gradient elutionfrom 8% B to 60% B in 60 min., detection 254 nm]. Fractions containingthe desired product were combined and lyophilized to providetrifluoroacetate salt of target product 132-1 (32.4 mg) as a whitesolid. LC-MS [M+H] 421.1.

Scale Yield Yield Purity Retention time Compound (mmol)* (mg) (%)* (%)[M + H] (min)** 132-1 0.15 32.4 40.5 97.9 421.1 5.63 *Based on theamount of compound 4 **HPLC-MS Method BEach of the following compounds was synthesized as described above using(S)-DAP(N′-Boc)-OMe×HCl or (S)-Me-DAP(N′-Boc)-OMe×HCl and theappropriate amine.

Ret. MH⁺ Time HPLC-MS Compound # Structure (m/z) (min) Method 132-2

435.1 4.04 A 132-3

485.1 6.55 B 132-4

485.1 4.49 A 132-5

499.1 4.60 A 132-6

499.1 4.61 A

Example 133N-((1S,2S)-2-Amino-1-hydroxycarbamoyl-propyl)-4-phenylethynyl-benzamide(133-1)

Synthesis of 4-phenylethynyl-benzoic acid (1)

The 4-ethynyl-benzoic acid (146 mg, 1.0 mmol), bromobenzene (116 μl, 1.1mmol), PdCl₂(dppf)₂ (24.5 mg, 0.03 mmol) and copper(I) iodide (11 mg,0.06 mmol), and diisopropylamine (7.0 mL, 2.3 mmol) were dissolved inTHF (2 mL) and sealed under nitrogen. The reaction mixture was thenstirred rapidly for 1 min and placed in the microwave reactor at 130° C.for 15 min. Then, mixture was diluted with EtOAc (50 mL) and water (40mL). After mixing and adjusting to pH˜5 with 3 M HCl, the aqueous phasewas removed. The organic phase was washed with 0.3 M HCl (20 mL×2), andbrine (30 mL). Filtration of the brown solution through anh Na₂SO₄ andevaporation of the solvents yielded the target crude product, which waspurified by HPLC purification. [Phenomenex Gemini C-18 column, 110 Å(30×100 mm); flow rate=20 mL/min; mobile phase A: 100% water, 0.1% TFA;mobile phase B: 100% ACN, 0.1% TFA; gradient elution from 30% B to 70% Bin 60 min., detection 254 nm]. Fractions containing the desired productwere combined and ACN was concentrated in vacuo. Product was extractedwith EtOAc (50 mL), washed with brine (50 mL×2), dried over anh. Na₂SO₄and solvent was evaporated in vacuo. Residue was dried in vacuoovernight to provide target compound 2 (160 mg, 72%) as white solid.LC-MS [M+H] 223.3.

Synthesis of(2S,3S)-3-(9H-fluoren-9-ylmethoxycarbonylamino)-2-(4-phenylethynyl-benzoylamino)-butyricacid methyl ester (2)

Reagent MW Eq. g/mL mmol Compound 1 222.25 1.0 53 mg 0.24H-(Allo)-Me-DAP-(N′-Fmoc)-OMe × 390.86 1.2 113 mg 0.29 HCl* HATU 380.231.2 110 mg 0.29 DIEA 129.24 3 125 μl 0.72 DMF 1.5 mL *amino acid wassynthesized according to Scheme 2

Compound 2 was prepared by reaction withH-(Allo)-Me-DAP-(N′-Fmoc)-OMe×HCl according to General Method for HATUcoupling in quantitative yield. LC-MS [M+H] 559.5 (C₃₅H₃₀N₂O₅+H,requires 559.64). Compound was used in next synthetic step withoutadditional purification.

Synthesis ofN-((1S,2S)-2-amino-1-hydroxycarbamoyl-propyl)-4-phenylethynyl-benzamide(133-1)

Reagent MW Eq. mg/mL mmol Compound 2 558.64 1.0 134 mg 0.24 NH₂OH × HCl69.49 10 167 mg 2.40 25% NaOMe/MeOH 12 658 μl 2.88 MeOH 3 mL THF 1.2 mL

Compound 133-1 was prepared by following General Method for hydroxamateformation, anhydrous. Crude compound was purified by HPLC. [PhenomenexGemini C-18 column, 110 Å (30×100 mm); flow rate=20 mL/min; mobile phaseA: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1% TFA; gradientelution from 15% B to 40% B over 60 min., detection 254 nm]. Fractionscontaining the desired product were combined and lyophilized to providethe trifluoroacetate salt of target compound 133-1 (21.1 mg) as a whitesolid. LC-MS: [M+H] 338.3.

Scale Yield Yield Purity Retention time Compound (mmol)* (mg) (%)* (%)[M + H] (min)* 133-1 0.24 21.1 mg 23.4 98.6 338.3 5.62 *Based on theamount of carboxylic acid used in the coupling reaction **HPLC-MS MethodBEach of the following compounds was synthesized as described abovestarting from appropriate carboxylic acid.

Ret. HPLC- MH⁺ Time MS Compound # Structure (m/z) (min) Method 133-2

437.1 2.88 B 133-3

407.1 3.24 B 133-4

368.3 3.41 BThe following compound was synthesized as described for synthesis ofcompound 132-1 in Example 132 using H-(Allo)-Me-DAP-(N′-Fmoc)-OMe×HCland Fmoc deprotection (20% piperidine in EtOAc).

Ret. HPLC- MH⁺ Time MS Compound # Structure (m/z) (min) Method 133-5

453.1 5.89 BEach of the following compounds was synthesized as described abovestarting from appropriate carboxylic acid.

Ret. MH⁺ Time HPLC-MS Compound # Structure (m/z) (min) Method 133-6 

352.3 5.68 B 133-7 

339 133-8 

338 133-9 

338 133-10

408.1 133-11

423.1 133-12

355.1 133-13

340.1 131-14

353.1 131-15

354.1

Example 134N—((S)-2-Amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-(2-phenyl-cyclopropylethynyl)-benzamide(134-1)

(2-phenylcyclopropyl)methanol (S2)

3.5 g Me₃SiCl (32 mmol) was added dropwise to the mixture of 53 g Zn/Cupowder and 50 ml Et₂O at stirring under N₂. The mixture was stirred for10 min and the solution of 1.6 ml CH₂I₂ and 10 ml Et₂O was added. Themixture was warmed to reflux and the solution of 26.7 ml CH₂I₂ (320mmol), 21.5 g ACHL025-1 (160 mmol) and 160 ml Et₂O was added dropwise atrefluxing. After the addition, the solution of 13.6 ml CH₂I₂ (160 mmol)and 30 ml Et₂O was added. The reaction mixture was stirred for 15 h.After the reaction, 300 ml Sat. NH₄Cl was added and the mixture wasfiltered. The organic layer was washed with Sat. NH₄Cl, brine, dried byMgSO₄, concentrated. The residue was purified by column chromatographyto give 9.7 g ACHL025-2 (41% yield). 1H-NMR (400 MHz, CDCl₃):

ppm: 0.91-0.99 (m, 2H), 1.43 (m, 1H), 1.81 (m, 1H), 3.52-3.62 (m, 2H),4.78 (s, 1H), 7.07-7.28 (m, 5H).

2-phenylcyclopropanecarbaldehyde (S3)

18.7 g IBX (66.9 mmol, 3.0 eq.) was added to the solution of 3.3 gACHL025-2 (22.3 mmol) in 60 ml EA. The mixture was warmed to 80° C.overnight. TLC showed the material was consumed completely. The reactionmixture was filtered, and the filtrate was concentrated to give 3.5 gcrude ACHL025-3. ¹H-NMR (400 MHz, DMSO):

ppm, 1.53-1.56 (m, 1H), 1.66-1.71 (m, 1H), 2.08-2.10 (m, 1H), 2.63-2.68(m, 1H), 7.16-7.29 (m, 5H), 9.07 (s, 1H).

(2-ethynylcyclopropyl)benzene (S4)

To a solution of diazophoaphonate (9.56 g, 50 mmol) and ACHL015-3 (8 g,54.8 mmol, 1.1 eq) in MeOH (200 ml) was added K₂CO₃ (13.8 g, 100 mmol).The mixture was stirred overnight at rt. The mixture was filtered andthe filtrate was concentrated to give crude. The crude was purified bycolumn chromatography on silica gel (PE/EA=50/0-50/1) to give 4.1 gACHL025-4 (57% yield). 1H-NMR (400 MHz, DMSO):

ppm, 1.23 (m, 2H), 1.59 (m, 1H), 2.21 (m, 1H), 2.79 (s, 1H), 7.10-7.26(m, 5H)

Methyl 4-((2-phenylcyclopropyl)ethynyl)benzoate (S5)

To a solution of 4.1 g ACHL025-4 (28.9 mmol) and 6.2 g methyl4-bromobenzoate (28.9 mmol) in 100 ml TEA was added 2 g PdC₂I (PPh₃)(2.89 mmol) and 0.55 g CuI (2.89 mmol) under N₂. The mixture was warmedto 90° C. and stirred overnighte. The reaction mixture was filtered andconcentrated to crude product. The crude product was purified by columnchromatography (PE/EA=100/0-100-1) to give 3.4 g (S5) (42% yield).¹H-NMR (400 MHz, CDCl₃):

ppm, 1.40-1.47 (m, 2H), 1.72-1.74 (m, 1H), 2.39-2.42 (m, 1H), 3.92 (s,3H), 7.13-7.32 (m, 5H), 7.45-7.47 (d, J=8 Hz, 2H), 0.7.96-7.98 (d, J=8Hz, 2H).

Methyl 4-((2-phenylcyclopropyl)ethynyl)benzoate (1)

To a solution of 4.1 g ACHL025-5 (14.8 mmol) in 30 ml MeOH and 20 mlwater was added 6 g NaOH. The mixture was stirred at rt for 4 h. 200 mlwater was added and the mixture was adjusted to PH=4 with 1N HCl. Themixture was extracted with EA and the organic layer was washed by brine,dried by MgSO₄, concentrated to give 3.3 g ACHL025 (85% yield). ¹H-NMR(400 MHz, DMSO):

ppm, 1.43 (m, 2H), 1.89 (m, 1H), 2.43 (m, 1H), 7.17-7.31 (m, 5H),7.50-7.52 (d, J=8 Hz, 2H), 7.90-7.92 (d, J=8 Hz, 2H), 13.10 (s, 1H)

Synthesis of(S)-3-tert-butoxycarbonylamino-3-methyl-2-[4-(2-phenyl-cyclopropylethynyl)-benzoylamino]-butyricacid methyl ester (2)

Reagent MW Eq. mg/mL mmol Compound 1 262.3 1.0 150 mg 0.57H—(S)-diMe-DAP-(N′-Boc)-OMe 246.3 1.15 162 mg 0.66 HATU 380.23 1.2 259mg 0.68 DIEA 129.24 3 298 μl 1.71 DMF 3 mL

Compound 2 was prepared by coupling with H—(S)-diMe-DAP-(N′-Boc)-OMeaccording to General Method for HATU coupling in quantitative yield.LC-MS [M+H] 492.0 (C₂₉H₃₄N₂O₅+H, requires 491.60). Compound was used innext synthetic step without additional purification.

Synthesis of(S)-3-amino-3-methyl-2-[4-(2-phenyl-cyclopropylethynyl)-benzoylamino]-butyricacid methyl ester (3)

Reagent MW Eq. mg/mL mmol Compound 2 490.6 1.0 280 mg 0.57 4NHCl/dioxane 14.0 2 mL 8.0 dioxane 2 mL

Compound 3 was prepared using the General Method for Boc deprotection inquantitative yield. LC-MS [M+H] 392.0 (C₂₄H₂₆N₂O₃+H, requires 391.49).Compound was used in next synthetic step without additionalpurification.

Synthesis ofN—((S)-2-amino-1-hydroxycarbamoyl-2-methyl-propyl)-4-(2-phenyl-cyclopropylethynyl)-benzamide(134-1)

Reagent MW Eq. mg/mL mmol Compound 3 × HCl 426.94 1.0 243 mg 0.57 50% aqNH₂OH 33 57.5 2 mL 32.8 i-PrOH 2 mL

Compound 134-1 was prepared by following General Method for hydroxamateformation, aqueous. Crude compound was purified by HPLC. [PhenomenexGemini C-18 column, 110 Å (30×100 mm); flow rate=20 mL/min; mobile phaseA: 100% water, 0.1% TFA; mobile phase B: 100% ACN, 0.1% TFA; gradientelution from 15% B to 60% B over 60 min., detection 254 nm]. Fractionscontaining the desired product were combined and lyophilized to providethe trifluoroacetate salt of target compound 134-1 (63.2 mg) as a whitesolid. LC-MS: [M+H] 392.3.

Scale Yield Yield Purity Retention Compound (mmol)* (mg) (%)* (%) [M +H] time (min)* 134-1 0.57 63.2 22.0 98.8 392.3 6.45 *Based on the amountof carboxylic acid used in the coupling reaction **HPLC-MS Method BThe following compound was synthesized as described above usingH—(S)-Me-DAP-(N′-Boc)-OMe.

Ret. HPLC- MH⁺ Time MS Compound # Structure (m/z) (min) Method 134-2

378.3 6.24 B

Example 135 Synthesis ofN-(1-(1-aminocyclopropyl)-2-(hydroxyamino)-2-oxoethyl)-4-((4-(morpholinomethyl)phenyl)ethynyl)benzamide(135-1)

Synthesis of 1-(2,2-dimethoxyethyl)cyclopropanol (2)

A solution of methyl 3,3-dimethoxypropanoate (28.2 g, 190 mmol) in athree-neck flask equipped with a overhead stirrer was cooled to 0° C.under N₂. The Ti(OiPr)₄ (11.3 mL, 38.1 mmol) was then slowly added andthe solution was allowed to stir for 10 min at 0° C. To this cooledsolution with vigorous stirring was then added the EtMgBr solution (158mL, 475 mmol) via syringe pump at ˜3 mL/min. After the addition wascomplete, the solution was allowed to slowly warm to ambient temperatureand stirred for an additional 18 h. The solution was then cooled to 0°C. and deionized water (45 mL) was slowly added with vigorous stirring.After stirring for 20 min at 0° C. the mixture vacuum filtered through apad of anhydrous MgSO₄. The clear, colorless solution was theconcentrated in vacuo and purified by flash chromatography (0-50%EtOAc/Hex) to provide the desired product (2) as a clear, colorless oil(8.60 g, 31%). ¹H NMR (DMSO) δ 5.04 (s, 1H), 4.62 (t, J=5.3, 1H), 3.22(s, 6H), 1.69 (d, J=5.3, 2H) 0.50 (dd, J=4.9, 6.9, 2H), 0.37 (dd, J=4.2,6.2, 2H).

Synthesis of 1-(2,2-dimethoxyethyl)cyclopropyl mesylate (3)

To a solution of compound 2 (8.60 g, 58.9 mmol) in CH₂Cl₂ was added TEA(12.6 mL, 124 mmol) sequentially. The solution was then cooled to 0° C.and methane sulfonyl chloride (8.09 g, 70.65 mmol) was added drop wise.After the addition was complete, the solution was stirred at 0° C. for10 min then allowed to warm to ambient temperature and stirred for 4 h.The solution was then washed with aqueous NaHSO₄ (1.0 N, 2×100 mL),aqueous NaHCO₃ (sat., 2×100 mL), dried (MgSO₄), filtered andconcentrated in vacuo. Isolated the desired product, (3) as a clear,colorless oil (13.62 g).

Synthesis of 2-(1-(methylsulfonyloxy)cyclopropyl)acetic acid (4)

Compound 3 (13.62 g, 60.8 mmol) was dissolved in THF (150 mL) and water(50 mL) and cooled to 0° C. To this cooled solution was added Oxone®(54.07 g, 88 mmol) in portions. After complete addition the slurry wasallowed to stir at 0° C. for 10 min the warmed to ambient temperature.After 8 h at ambient temperature the slurry was diluted with deionizedH₂O (100 mL) and extracted with EtOAc (3×150 mL). The combined organicswere dried (MgSO₄), filtered and concentrated in vacuo. The product (4)was isolated as a thick, colorless oil (8.4 g).

Synthesis of methyl 2-chloro-2-cyclopropylideneacetate (5)

Compound 4 (4 g, 20.6 mmol) was dissolved in anhydrous DCE (100 ml)under N₂ and cooled to 0° C. Thionyl chloride (1.8 ml, 30 mmol) was thenadded slowly and the solution was refluxed for 30 min. At ambienttemperature, were added NCS (3.32 g, 25 mmol) and 4 drops ofconcentrated HCl and the solution was heated under reflux for 8 h. MeOH(50 ml) was added at room temperature and the solution was stirred at rtfor 3 h, Followed by TEA (10 ml. 100 mmol) and stirring was continuedfor additional 1 h The solvent were removed under reduced pressure,water (100 ml) was added and product was extracted with ethyl acetate,dried (Na2SO4), and and purified by flash chromatography (0-15% EA inHex) to give 1.2 g of methyl 2-chloro-2-cyclopropylideneacetate 5. H1NMR (DMSOd6) 1.5 (t, 2H), 1.73 (t, 2H) 3.73 (s, 3H)

Synthesis of methyl2-chloro-2-(1-(diphenylmethyleneamino)cyclopropyl)acetate (6)

To diphenyl imine (2.05 g, 11.37 mmol) in methanol (35 ml), the chlorideproduct 5 (1.39 g, 9.48 mmol) was added in dry methanol (14 ml) andstirred at room temp for 24 h. The reaction mixture was concentrated togive 3.39 g of 6:1H NMR (CDCl3) 0.70-0.9 (m, 3H), 1.20-1.24 (m, 1H) 3.62(s, 3H), 4.29, (s, 1H), 7.23-7.58 (m, 10H).

Synthesis of methyl2-azido-2-(1-(diphenylmethyleneamino)cyclopropyl)acetate (7)

The chloride product 6 (3.24 g, 99 mmol) was redessolved in dry DMF (25ml), NaN3 (1.3 g, 200 mmol) was added and heated at 85° C. for 18 h. Thereaction mixture was diluted with ethyl acetate (100 ml) and water 5 mlwas added. The organic layer was separated, dried and concentrated togive 2.8 g of 7. LCMS (M+1) 335, Chemical Formula: C19H18N4O2 MW:334.37.

Synthesis of methyl2-amino-2-(1-(diphenylmethyleneamino)cyclopropyl)acetate (8)

The compound 7 (480 mg, 1.43 mmol), PPh3 (278 mg, 1.06 mmol) in THF (5ml) and water (1 ml) stirred for 18 h. The reaction mixture was dilutedwith ethyl acetate (100 ml) and water 5 ml was added. The organic layerwas separated, dried and concentrated to give crude 8 which were usedfor the next step without any further purification.

Synthesis of methyl2-(4-(cyclopropylbuta-1,3-diynyl)benzamido)-2-(1-hydroxycyclopropyl)acetate(9)

A solution of compound 4-((4-(morpholinomethyl)phenyl) ethynyl)benzoicacid (100 mg, 0.31 mmol), HATU (120 mg, 0.31 mmol) and DIEA (0.5 ml,excess) in CH3CN (15 mL) was maintained at ambient temperature for 10min followed by the addition of compound 8 (408 mg, 1.31 mmol). Reactionmixture was stirred at ambient temperature 20 min and concentratedfollowed by the dilution with water (10 ml). Solution was extracted withethyl acetate (20 ml×2) and brine (20 ml). Organic layer was dried overMgSO₄ and evaporated to give crude product 9, 160 mgs of methyl2-(1-aminocyclopropyl)-2-(4-((4-(morpholinomethyl)phenyl)ethynyl)benzamido)acetate. LC-Ms (M+1) 448; Chemical Formula: C26H29N3O4; MW:447.53.

Synthesis ofN-(1-(1-aminocyclopropyl)-2-(hydroxyamino)-2-oxoethyl)-4-((4-(morpholinomethyl)phenyl)ethynyl)benzamide(135-1)

Aq. Hydroxylamine (2 ml, 50% aq.) was added to a stirred solution ofester 9, (0.16 g, 0.35 mmol) in isopropanol (15 ml), stirred for 18 h.Excess solvent was removed and the product was purified on a reversephase HPLC to give compound (135-1) (6 mg). LC-MS (M+1) 449: ChemicalFormula: C25H28N4O4; MW: 448.51.

Example 136N-((2S,3R)-3-amino-1-(hydroxyamino)-1-oxobutan-2-yl)-4-(4-(morpholinomethyl)phenethyl)benzamide(137-1)

Compound 1 (200 mg, 0.45 mmol) was dissolved in methanol (20 ml)followed by the addition of Pd/C (10% wt, 46 mg). Reaction mixture wassubjected to hydrogenation (Parr apparatus, 46 psi) at room temperaturefor 15 min. Catalyst was filtered and washed with methanol. Filtrate wasevaporated in vacuum to provide target compound 136 (184 mg) ascolorless oil. LC-MS [M+H] 441.0 (C10H20N2O4+H, requires 440.24).

Example 137

The following compounds were made according to the above syntheticprocedures.

Compound MH⁺ # Structure (m/z) 137-1 

383.4 137-2 

534.6 137-3 

465.5 137-4 

381.4 137-5 

382.4 137-6 

421.5 137-7 

445.5 137-8 

442.5 137-9 

358.4 137-10

370.4 137-11

452.4 137-12

365.4 137-13

441.5 137-14

383.4 137-15

341.4 137-16

449.4 137-17

467.5 137-18

398.4 137-19

364.4 137-20

368.4 137-21

302.3 137-22

369.4 137-23

383.5 137-24

450.5 137-25

344.4 137-26

364.4 137-27

364.4

Example 138

The following compounds may be made according to the above syntheticprocedures.

Compound MH⁺ # Structure (m/z) 138-1

460.5 138-2

461.6 138-3

363.4 138-4

300.3 138-5

324.3 138-6

473.6 138-7

329.4 138-8

343.3 138-9

338.4  138-10

344.4  138-11

437.5  138-12

437.5

LC-MS Analytical Methods

All LC-MS spectra were acquired using HPLC (Column: Phenomenex, OnyxMonolithic C18, 4.6×50 mm. Flow rate: 1.5 mL/min. Mobile phase A: 0.1%trifluoroacetic acid in water. Mobile phase B: 0.1% trifluoroacetic acidin acetonitrile.) with an ion spray MS detector (PE SCIEX API 365LC/MS/MS), and UV/VIS (254 nm) detection (Shimadzu SPD-10A) under thefollowing gradients:

Method A (12 minute method): 5% B to 100% B over 9.6 min.

Method B (Analytical 5-60): 5% B to 60% B over 9.1 min.

Method C (6 minute method): 5% B to 100% B over 4.6 min.

Method D (Analytical 5-35): 5% B to 35% B over 9.1 min.

NMR Data

All NMR spectra were taken on a Bruker 250 NMR (250 MHz) or Varian 400(400 MHz), using DMSO-d₆ or CDCl₃

Biological Protocols and Data Bacterial Screens and Cultures

Bacterial isolates were cultivated from −70° C. frozen stocks byovernight passages at 35° C. in ambient air on Mueller-Hinton agar(Beckton Dickinson, Franklin Lakes, N.J.). Clinical isolates tested wereobtained from various geographically diverse hospitals in the US andabroad (Focus Diagnostics, Herndon, Va. and JMI, North Liberty, Iowa).Quality control strains were from the American Type Culture Collection(ATCC; Rockville, Md.). Additional primary panel strains were generatedusing standard molecular biology techniques. Strain AECO063 does notexpress the principal multidrug efflux pump for E. coli and ishypersusceptible to many antibacterials. Target enzyme replacementstrains (AECO061 —E. coli, AECO062 —P. aeruginosa and AECO065 —A.baumannii) possess a non-functional genomic copy of the E. coli lpxCgene complemented by a plasmid-borne sequence from the species indicatedunder the control of the native E. coli lpxC promoter (E. coli strainbackground MG1655).

Susceptibility Testing

Minimum Inhibitory Concentrations (MICs) were determined by the brothmicrodilution method in accordance with the Clinical and LaboratoryStandards Institute (CLSI) guidelines. In brief, organism suspensionswere adjusted to a 0.5 McFarland standard to yield a final inoculumbetween 3×10⁵ and 7×10⁵ colony-forming units (CFU)/mL. Drug dilutionsand inocula were made in sterile, cation adjusted Mueller-Hinton Broth(Beckton Dickinson). An inoculum volume of 100 μL was added to wellscontaining 100 μL of broth with 2-fold serial dilutions of drug. Allinoculated microdilution trays were incubated in ambient air at 35° C.for 18-24 hours. Following incubation, the lowest concentration of thedrug that prevented visible growth (OD600 nm<0.05) was recorded as theMIC. Performance of the assay was monitored by the use of laboratoryquality-control strains and levofloxacin, a compound with a defined MICspectrum, in accordance with CLSI guidelines. Typically, compounds ofthe present invention have MIC values of 0.03-64 mic/mL. To this end,data for certain representative compounds is shown in Table 1 below.

Efficacy in Mouse Model of Systemic Pseudomonas aeruginosa andEscherichia coli Infection

Male CD-1 mice weighing 22-26 g were infected intraperitoneally with 0.5mL of a bacterial suspension of P. aeruginosa strain ATCC27853 or E.coli ATCC25922 in blood heart infusion broth (BHI, DIFCO, USA) plus 5%Mucin (Sigma, USA). The number of bacterial cells used was approximately2 times the dose that would kill 90% of animals (LD₉₀, 6.5×10⁴ CFU forP. aeruginosa, 4.5×10⁵ CFU for E. coli). At one hour (1 hr) postinfection, the test compound was injected intravenously in doses rangingfrom 0.3 mg/kg to 50 mg/kg, ten mice per dosing group. Survival of themice was observed daily for 7 days, and the dose of compound resultingin survival of 50% of mice (ED₅₀) was calculated. Example 3/12ademonstrated an ED50<20 mg/kg in the E. coli septicemia model.

Efficacy in Neutropenic Thigh Model

Mice were rendered neutropenic prior to infection with 2 doses ofcyclophosphamide, and then infected intramuscularly in the thigh with10⁵-10⁶ CFU of bacteria. See Gudmundsson S., Erlensdottir H., “MurineThigh Infection Model” Handbook of Animal Models of Infection, M. A.Sande and O. Zak, Eds.; London: Academic Press, 1999, pp 137-144.Antibiotics or vehicle alone as a negative control were administeredtwice at 2 hrs and 14 hrs post-infection. At 24 hrs post-infection,thighs were harvested, homogenized, and plated to measure the number ofCFUs surviving per thigh. Thighs from a subset of animals were alsoharvested 2 hrs post-infection to record the CFUs present just prior tothe first antibiotic treatment (pre-treatment). The static dose, definedas the dose required to result in a CFU load at 24 hours that isidentical to that measured at 0 hours post infection, was calculated bystandard methods in Prizm from a dose response curve.

TABLE 1 AECO 001 APAE Compound AECO HSA AECO APAE APAE 006 + # 001 AGP063 001 006 PMBN AABA1060 ASAU001  62-6 C D B D  62-7 C C B D B C D 62-10 B D A D  62-8 C D C D  62-11 B D A D  62-9 C D B D  62-4 B D B D 62-5 B D C D  48-5 D D D D D D D  47-5 D D D D C D D  46-4 C C B C A DD  41-5 D D C C A A D D  43-7 D D C C A A D D  42-7 C C C C A A C D 60-5 C D C D C B D D 115-2 A B A A A A B D 115-3 A A A B A A B D  66-5A B A B A A B D  67-9 B C A C A A D D  67-7 A B A B A A B D  67-8 A B AB A A B D  65-6 C D B C B A C D  64-7 B C A B A A C D  59-8 C D C D C CD D  67-5 A B A B A A C D  68-6 A B A B A A C D  66-6 A B A C A A C D 30-12a A A A A A A B D  30-12b B C A B A A C D  30-11a B C B C B A C D115-4 B C A A A A C D 115-5 A B A B A A B D 115-6 B B A A A A C D 115-7A B A B A A B D 115-8 A B A A A A B D  96-1 A A A B A A B D 110-3 A B AA A A B D 110-6 A B A B A A B D 111-3 A B A B A A B D 110-7 A B A B A AA D 111-2 B B A B A A B C 111-4 A C A B A A C D 110-4 A B A B A A C C110-2 A B A B A A B D 118-10 A B A C B A B D  96-4 A B A B A A B D 132-4A B A B A A B C 113-2 A B A A A A B D 113-1 A B A B A A C D 113-3 A B AB A A B D 113-4 A B A B A A B D 110-5 A B A B A A B D 112-1 A C A A A AB D 112-2 B C A B A A D D 112-3 A C A B A A C C 110-8 A C A B A A C D110-9 A B A B A A B D 110-10 A B A B A A C C 110-11 A B A C A A C C110-12 A A A B A A A C 112-4 A B A A A A A C 112-5 A B A A A A A C 127-1C C B D C C D D  97-1 A A A B A A B C  97-2 A A A A A A A C  98-1 A A AA A A C C 116-1 C C C B A A C C 117-1 D D D D D D D D 117-2 D D D D D CD D 118-1 C C B B A A C C 118-2 A C A A A A B C 118-3 A B A B A A B C128-1 C C C C C C C C 113-11 A C A B A A C D  96-2 A B A B A A B C  96-3A B A B A A B C  98-3 A A A B A A C D  98-2 A B A B A A C C  99-1 A B AB A A C C 100-1 A B A B A A C C 119-1 A B A A A A B C 119-2 B C B A A AD D 119-3 B B A B A A C D 131-1 C D C B A A D D 131-2 B C A A A A C D131-3 C C C B A A C C  82-2 A C A A A A C C  82-3 A C A A A A B C  86-2A A A B A A B C  82-4 A B A B A A B B 101-2 A C A B A A C C 102-3 A B AA A A C C 101-3 A B A A A A B C 101-1 A B A B A A C C 101-4 A B A B A AC C 102-2 A B A B A A B C 102-1 A B A B A A C C 102-4 A B A B A A C D133-7 A B A A A A A B 133-8 C C B B A A C D 133-9 C C B B A A C D  86-3A A A A A A B C  82-5 A B A A A A B B  82-6 A B A B B A C C 113-9 A B AA A A A C 113-10 A B A B A A B C 103-1 A B A B A A D D 137-1 A B A B A AD D 137-2 A B A B A A C D 137-3 A B A B A A C C 120-3 A B A A A A B D113-8 A B A B A A A C 104-2 A B A B A A C C 104-1 A B A B A A B C 104-3A B A A A A A C 104-4 A B A A A A B C 113-7 A B A B A A B D 113-5 A A AA A A A C 101-5 A B A A A A A C 137-4 D D C D C C D D  82-7 A B A A A AA B  82-8 A B A C A A D D  82-9 A B A B A A B B  82-10 A B A B A A B B 82-11 B B A C B A C C  88-3 A B A A A A B D  88-4 B C A A A A C D  91-2A B A A A A A B  82-12 A B A B A A B C 102-5 A A A A A A B D  88-2 A B AA A A B B 133-2 A C A A A A C D 109-1 A B A B A A B D 137-5 A B A A A AB C 129-1 A B A A A A B D 137-6 A B A A A A B C 137-7 A B A A A A B C120-1 A B A A A A B C  82-13 A A A A A A A C 133-4 B C A A A A D D 136-1B C A B A A C C 137-8 B C A B A A C C 137-9 A B A B A A B C 118-4 A B AA A A C D 118-5 B B A A A A C D 118-6 B B A A A A C C 118-7 A B A A A AB C  85-2 A B A B A A C C  85-1 A B A B A A C C  87-2 A A A A A A C C 87-3 A A A A A A A C 133-3 A B A A A A C D 133-1 A B A A A A A C 133-10B B A C A A C C 133-11 A C A C B A C C 118-8 A B A A A A B C  88-1 A B AB A A C C 133-5 A B A C A A A C 114-2 A B A B A A B C 105-1 A B A B A AC C 105-3 C C B C B A C C 109-2 A B A B A A B C 133-12 A B A A A A C C118-9 A B A A A A B C 119-6 B B A A A A C C 114-3 A B A A A A A C 114-4A B A B A A B B 114-5 A B A B A A A B 114-6 A B A B B A B B 114-7 B B AB B A B B 114-8 A B A A A A B C 114-9 B B A A A A B C 114-1 A B A A A AA B  88-9 A B A B A A C C  88-10 A B A B A A B C 114-10 A B A B A A B C137-26 C C B C B A C C 137-27 C C B C A A C C  82-14 A C A A A A A C 82-15 A C A A A A A C  88-5 A A A B A N/A A C 137-12 B C B B A A C C 91-3 A B A A A A B B  91-4 A B A B A A B B 120-2 B C A B A A C C  82-16A B A A A A A C 105-2 A A A B A A C C 105-4 A B A C B A C C 105-5 B B AC B A C C  91-5 A A A A A A B C  82-17 A A A A A A A B 119-4 A A A A A AB C 119-5 A A A A A A A C 133-13 C C B C A A C C 133-15 B B A B A A B C133-14 A A A B A A A C 137-13 C C B C A A C C  91-30 C C B A A A C C130-1 B C A A A A B C  82-18 A B A B A A C C  82-19 A B A B A A A C 91-6 A A A A A A A C 114-11 A C A C A A B C 137-14 A C A B A A B C114-12 A B A B A A A C 114-13 A B A B A A B C 114-14 A B A B A A A C 82-20 A B A B A A B C  82-21 A B A B A A A C 132-1 A B A B A A B C132-3 A B A C A A B C  88-12 A B A A A A A B  92-2 C C B C B B C C 91-17 A B A A A A B C  82-22 A C A C C A C C  91-10 A A A A A A A C 87-1 A B A A A A A C  88-6 A B A B A A B C 104-5 A A A B A A A B 126-2A A A B A A A C 121-1 A A A A A A B C 121-2 B C B C A A C C 101-6 A C AA A A B C  92-1 B C B B A A C C 122-1 B B A B A A C C 123-1 A B A B A AB C 137-16 A B A B A A B C 106-6 A C A B A A B C 106-7 A B A B A A B C137-17 C C C C B A C C 137-18 C C C C B B C C 137-19 A B A B A A C C124-1 A B A B A A C C  91-11 A C A B A A C C  88-7 A B A B A A B C 91-12 A A A A A A A B 124-2 A B A B A A B C 124-3 A B A B A A B C 132-5C C N/A C A N/A C C 132-2 A B A B A A C C 132-6 B C A C A A C C 106-4 BC A B A A C C  91-13 A B A A A A B B  82-23 A C A C A A C C 114-15 B C AA A A B C  86-4 A B A B A A B C  82-24 A B A B A A A B 107-2 A B A A A AA B 106-1 A B A C A A C C 107-3 A B A A A A B C 106-5 A B A B A A B C107-1 A B A A A A B C 106-3 A C A A A A C C 137-20 C C C C B A B C 133-6A A A A A A A B  91-14 A B A A A A A B  82-25 A A A A A A A C  82-26 A CA B B A B B  91-1 A A A A A A A B 106-2 A B A B A A B C  91-7 A B A A AA B B  91-8 A B A A A A B B  89-1 A B A A A A B C  93-1 A C A A A A A C137-21 C C C B A A B C 114-16 A A A B A A A C 107-4 A A A A A A A B122-5 A A A A A A A C  88-8 A A A A A A B C 122-6 A A A A A A B C  91-15A B A A A A A B 125-1 A B A B A A A C  86-1 A B A A A A B C 137-22 C C CB B B C C 137-23 C C C B A B C C 137-24 A B A B A A B C  94-1 A B A A AA B C  82-1 A B A B A A B C  84-1 A B A C B A C C  86-5 A C A A A A B C114-17 A B A B A A B B  91-9 A A A A A A A B 137-25 C C B C A A C C107-5 A B A B A A A B  88-13 A A A A A A A B  88-15 A A A A A A A B 83-2 A C A A A A B C  91-16 A A A A A A A B  83-1 A B A A A A A C 88-11 A A A A A N/A A C  88-14 A B A A A A A C 107-6 A B A B A A A C134-1 B B A B A A A C 134-2 B C B B A A B C 122-8 A A A A A A B C 126-1A A A A A A A C  82-27 A B A B A A B C  89-2 A B A A A A A C 114-18 A BA A A A A C 122-2 A B A A A A B C 122-3 A A A A A A A C 122-4 A B A A AA A C 102-6 A B A A A A B C  89-3 A A A A A A B C  91-21 A B A B A A B B 91-22 A B A B A A B B  91-23 A B A B A A B B 138-10 A B A B A A B B 87-4 A B A A A A A C  88-16 A B A A A A A C  89-4 C C B B A A C C  89-5B B A A A A B C  82-28 A B A B A A A B 113-6 A A A A A A A C  91-18 A AA B B A B C  91-19 A A A B A A B C  91-20 A B A B B A B C  89-6 B B A AA A B C  89-7 B B A B A A C C 114-19 A B A A A A A B 114-20 A B A B A AA B  82-29 A A N/A A A N/A A B  86-6 A B A B A A B C  86-7 A B A B A A AC  91-24 A B A A A N/A B B  91-25 C C C C B B C C  91-26 A A A A A N/A AC  91-27 A B A A A N/A B C  91-28 A B A B A A B B  88-17 A C A A A A B C115-1 A B A B A A B D 137-11 D D C C B B D D 137-10 D D D D C C D D MICKey MIC's of 2.0 μg/mL or less = A MIC's of greater than 2.0 μg/mL to16.0 μg/mL = B MIC's of greater than 16.0 μg/mL to 64.0 μg/mL = C MIC'sof greater than 64.0 μg/mL = D *AEC001 is ATCC25922, AECO063 isATCC25922 bearing an acrAB mutation, APAE001 is ATCC27853, APAE006 is astrain of P. aeruginosa in which all efflux pumps are inactivated,APAE006 + PMBN indicates MICs with the indicated strain in the presenceof 8 micrograms per milliliter of polymyxin B nonapeptide, AABA1060 is aclinical isolate of Acinetobacter baumannii, ASAU001 is a laboratorystrain of S. aureus (ATCC29213).

As shown in the following Table 2, the MICs on AECO001 for fourteenpairs of compounds were compared (n=2 data). For each pair of compounds,the compounds were identical in structure except for the A moiety. Morespecifically, A is —C(CH₃)₂NH₂ in one compound and A is —CHCH₃NH₂ in theother compound. As shown, for each pair of compounds, the compoundwherein A is —C(CH₃)₂NH₂ was more potent.

TABLE 2 Compound # Ratio of MICs on AECO001 (A = Compound # [Cpd (A =—CHCH₃NH₂)/ —CHCH₃NH₂) (A = —C(CH₃)₂NH₂) Cpd (A = —C(CH₃)₂NH₂)] 91-291-12 5.6 91-3 91-14 3.3  82-16 82-25 1.6 91-7 91-1  7.2 88-3 88-8  5.1 91-13 91-15 6.5 91-8 91-9  5.2  88-12 88-13 4.1  91-17 91-16 2.6 89-189-2  1.6 115-2  121-1  4.0 133-7  133-6  20.8 118-7  122-5  2.4 118-9 122-6  2.1

As shown in the following Table 3, three pairs of compounds were testedin the neutropenic thigh efficacy model. For each pair of compounds, thecompounds were identical in structure except for the A moiety. Morespecifically, A is —C(CH₃)₂NH₂ in one compound and A is —C(CH₃)₂OH inthe other compound. A ratio of >1 indicates that the compound wherein Ais —C(CH₃)₂NH₂ had a lower static dose and is therefore a more potentcompound in vivo. For all three pairs of compounds, the compound whereinA is —C(CH₃)₂NH₂ had superior in vivo efficacy to that of the compoundwherein A is —C(CH₃)₂OH. In addition, the ratios of the MICs of eachpair of compounds on the infecting strain (ATCC43816) was calculated. Aratio of >1 indicates that the compound wherein A is —C(CH₃)₂NH₂ had alower MIC and was therefore more potent on this strain. When the ratioof neutropenic thigh potencies was normalized to reflect the differencesin MICs, it was found that the compounds wherein A is —C(CH₃)₂NH₂ stillhave superior activity relative to the compounds wherein A is—C(CH₃)₂OH, demonstrating that the functional group —C(CH₃)₂NH₂ confersadvantageous properties for in vivo antibacterial activity that is notreadily predictable from the MICs.

TABLE 3 Ratio of Ratio of MICs on thigh static Ratio of ATCC43816 doseCompound # Compound # thigh static [Cpd (A = —CH(CH₃)₂OH/Cpd normalized(A = —C(CH₃)₂NH₂) (A = —C(CH₃)₂OH) doses (A = —C(CH₃)₂NH₂)] to MICs88-13 88-11 >7x 1.6 >4.4x 91-12 91-6  >3x 1.0 >3x 122-5  122-8  >2x 0.5>4x

It should be understood that the organic compounds according to theinvention may exhibit the phenomenon of tautomerism. As the chemicalstructures within this specification can only represent one of thepossible tautomeric forms, it should be understood that the inventionencompasses any tautomeric form of the drawn structure.

Furthermore, while particular embodiments of the present invention havebeen shown and described herein for purposes of illustration, it will beunderstood, of course, that the invention is not limited thereto sincemodifications may be made by persons skilled in the art, particularly inlight of the foregoing teachings, without deviating from the spirit andscope of the invention. Accordingly, the invention is not limited exceptas by the appended claims.

All of the U.S. patents, U.S. patent application publications, U.S.patent applications, foreign patents, foreign patent applications andnon-patent publications referred to in this specification areincorporated herein by reference, in their entirety to the extent notinconsistent with the present description.

What is claimed is:
 1. A compound having the following formula (I):

or a stereoisomer, pharmaceutically acceptable salt, ester, or prodrugthereof, wherein: E is selected from the group consisting of: (1) H, (2)substituted or unsubstituted C₁-C₆-alkyl, (3) substituted orunsubstituted C₂-C₆-alkenyl, (4) substituted or unsubstitutedC₂-C₆-alkynyl, (5) substituted or unsubstituted C₃-C₁₀-cycloalkyl, (6)substituted or unsubstituted aryl, (7) substituted or unsubstitutedheterocyclyl, and (8) substituted or unsubstituted heteroaryl; L isabsent or selected from the group consisting of: (1) substituted orunsubstituted C₁-C₆-alkyl, (2) —(NR^(3L))₀₋₁—(CH₂)₀₋₄—NR^(3L)—(CH₂)₀₋₄—,(3) —(NR^(3L))₀₋₁—C(R^(1L),R^(2L))—NR^(3L)—C(R^(1L),R^(2L))—, (4)—C(R^(1L),R^(2L))—O—C(R^(1L),R^(2L))—, (5)—(CH₂)₀₋₄—NR^(3L)—C(R^(1L),R^(2L))—CONH—(CH₂)₀₋₄—, (6)—CO—C(R^(1L),R^(2L))—NHCO—, (7) —CONR^(3L)—, (8) —NR^(3L)CO—, (9)—NR^(3L)—, (10) —SO₂NR^(3L)—, (11) —NR^(3L)—C(═O)—NR^(3L)—, (12)substituted or unsubstituted C₃-C₁₀-cycloalkyl, (13) substituted orunsubstituted aryl, (14) substituted or unsubstituted heterocyclyl, and(15) substituted or unsubstituted heteroaryl, wherein: each R^(1L),R^(2L), and R^(3L) is independently selected from the group consistingof:  (a) H,  (b) substituted or unsubstituted C₁-C₆-alkyl,  (c)C₁-C₆-alkyl substituted with aryl,  (d) C₁-C₆-alkyl substituted withheterocyclyl, and  (e) C₁-C₆-alkyl substituted with heteroaryl, orR^(1L) and R^(3L), together with the atoms to which they are attachedcan form a substituted or unsubstituted heterocyclic ring, having from 3to 8 ring atoms, wherein 1-2 ring atoms of the heterocyclic ring areselected from N, O and S; D is absent or selected from the groupconsisting of: (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl, (2)substituted or unsubstituted aryl, (3) substituted or unsubstitutedheterocyclyl, and (4) substituted or unsubstituted heteroaryl; G isselected from the group consisting of: (1) —NR^(1G)C(═O)—, (2)—C(═O)NR^(1G)—, (3) —(CH₂)₀₋₄NHCH₂C(═O)NR^(1G)—, (4) —CR^(2G)═CR^(2G)—,(5) —S(═O)—, (6) —SO₂—, (7) —C(R^(3G))₂—S(═O)—, (8) —S(═O)—C(R^(3G))₂—,(9) —C(R^(3G))₂—SO₂—, (10) —SO₂—C(R^(3G))₂—, (11)—CR^(3G)═CR^(3G)—CR^(3G)═CR^(3G)—, (12) —C(R^(3G))₂—, (13)—CR^(3G)═CR^(3G)—C≡C—, (14) —C≡C—CR^(3G)═CR^(3G)—, (15) —C(═O)—C≡C—,(16) —C≡C—C(═O)—, (17) substituted or unsubstituted C₃-C₁₀-cycloalkyl,(18) substituted or unsubstituted aryl, (19) substituted orunsubstituted heterocyclyl, and (20) substituted or unsubstitutedheteroaryl, wherein: R^(1G) is substituted or unsubstituted C₁-C₆-alkyl;each R^(2G) is independently selected from the group consisting of H, ahalogen atom, and substituted or unsubstituted C₁-C₆-alkyl, and at leastone R^(2G) is not H; and R^(3G) is selected from the group consisting ofH, a halogen atom, and substituted or unsubstituted C₁-C₆-alkyl; Y isabsent or selected from the group consisting of: (1) substituted orunsubstituted C₃-C₁₀-cycloalkyl, (2) substituted or unsubstituted aryl,(3) substituted or unsubstituted heterocyclyl, and (4) substituted orunsubstituted heteroaryl; X is selected from the group consisting of:(1) —(C═O)NR₄—, (2) —C₁-C₆-alkyl-(C═O)NR₄—, (3)—C₂-C₆-alkenyl-(C═O)NR₄—, (4) —C₂-C₆-alkynyl-(C═O)NR₄—, (5) —CH₂NR₄—,(6) —SO₂NR₄—, (7) —S(═O)NR₄—, (8) —NR₄C(═O)—, and (9) —NR₄—, or X and A,together with the atoms to which they are attached can form aheterocyclic ring, having from 5 to 8 ring atoms, wherein 1-2 ring atomsof the heterocyclic ring are selected from N, O and S, or when Y is abicyclic substituted or unsubstituted heterocyclyl or heteroaryl, then Xis absent; R₃ is H or substituted or unsubstituted C₁-C₆-alkyl, or R₃and A, together with the atom to which they are attached can form asubstituted or unsubstituted 3-10 membered cycloalkyl or a heterocyclicring, having from 3 to 10 ring atoms, wherein 1-4 ring atoms of theheterocyclic ring are selected from N, O and S; R₄ is (1) H orsubstituted or unsubstituted C₁-C₆-alkyl, or (2) R₄ and A, together withthe atoms to which they are attached can form a substituted orunsubstituted heterocyclic ring, having from 3 to 8 ring atoms, wherein1-2 ring atoms of the heterocyclic ring are selected from N, O and S, or(3) R₄ and Y, together with the atoms to which they are attached, form abicyclic substituted or unsubstituted heterocyclyl or heteroaryl; n isan integer from 0-6; A is selected from the group consisting of: (1) H,(2) —(CH₂)₀₋₄C(R^(1a),R^(2a))(CH₂)₀₋₄OR^(3a), (3)—(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a),R^(5a)), (4)—(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a))COR^(3a), (5)—(CH₂)₀₋₄C(R^(1a),R^(2a))NHCON(R^(4a),R^(5a)), (6)—(CH₂)₀₋₄C(R^(1a),R^(2a))NHC(═NH)N(R^(4a),R^(5a)), (7)—CH(R^(1a),R^(2a)), (8) —C≡CH, (9) —(CH₂)₀₋₄C(R^(1a),R^(2a))CN, (10)—(CH₂)₀₋₄C(R^(1a),R^(2a))CO₂R^(3a), (11)—(CH₂)₀₋₄C(R^(1a),R^(2a))CON(R^(4a),R^(5a)), (12) substituted orunsubstituted C₃-C₁₀-cycloalkyl, (13) substituted or unsubstituted aryl,(14) substituted or unsubstituted heterocyclyl, and (15) substituted orunsubstituted heteroaryl, wherein: each R^(1a), R^(2a), R^(3a), R^(4a),and R^(5a) is independently selected from the group consisting of:  (a)H,  (b) a halogen atom,  (c) substituted or unsubstituted C₁-C₆-alkyl, (d) substituted or unsubstituted aryl,  (e) substituted orunsubstituted heterocyclyl, and  (f) substituted or unsubstitutedheteroaryl, or R^(4a) and R^(5a) together with the N atom to which theyare attached can form a substituted or unsubstituted heterocyclic ring,having from 5 to 8 ring atoms, wherein 1-2 ring atoms of theheterocyclic ring are selected from N, O and S; Q is absent or selectedfrom the group consisting of: (1) —C(═O)N(R₁,R₂), (2) —NHC(═O)N(R₁,R₂),(3) —N(OH)C(═O)N(R₁,R₂), (4) —CH(OH)C(═O)N(R₁,R₂), (5) —CH[N(R^(2q),R^(3q))]C(═O)N(R₁,R₂), (6) —CHR^(1q)C(═O)N(R₁, R₂), (7) —CO₂H, (8)—C(═O)NHSO₂R^(4q), (9) —SO₂NH₂, (10) —N(OH)C(═O)R^(1q), (11)—N(OH)SO₂R^(4q), (12) —NHSO₂R^(4q), (13) —SH, (14)—CH(SH)(CH₂)₀₋₁C(═O)N(R₁,R₂), (15) —CH(SH)(CH₂)₀₋₁CO₂H, (16)—CH(OH)(CH₂)₀₋₁CO₂H, (17) —CH(SH)CH₂CO₂R^(1q), (18) —CH(OH)(CH₂)SO₂NH₂,(19) —CH(CH₂SH)NHCOR^(1q), (20) —CH(CH₂SH)NHSO₂R^(4q), (21)—CH(CH₂SR^(5q))CO₂H, (22) —CH(CH₂SH)NHSO₂NH₂, (23) —CH(CH₂OH)CO₂H, (24)—CH(CH₂OH)NHSO₂NH₂, (25) —C(═O)CH₂CO₂H, (26) —C(═O)(CH₂)₀₋₁CONH₂, (27)—OSO₂NHR^(5q), (28) —SO₂NHNH₂, (29) —P(═O)(═O)₂,

and (33) —N(OH)C(═O)CR₁R₂, wherein: R₁ is selected from the groupconsisting of:  (1) —H,  (2) —OH,  (3) —OC₁-C₆-alkyl,  (4) —N(R^(2q),R^(3q)), and  (5) substituted or unsubstituted C₁-C₆-alkyl; R₂ isselected from the group consisting of:  (1) H,  (2) substituted orunsubstituted C₁-C₆-alkyl,  (3) substituted or unsubstitutedC₂-C₆-alkenyl,  (4) substituted or unsubstituted C₂-C₆-alkenyl,  (5)substituted or unsubstituted aryl,  (6) substituted or unsubstitutedheterocyclyl, and  (7) substituted or unsubstituted heteroaryl, or R₁and R₂, together with the N atom to which they are attached can form asubstituted or unsubstituted heterocyclic ring, having from 3 to 10 ringatoms, wherein 1-4 ring atoms of the heterocyclic ring are selected fromN, O and S; and each R^(1q), R^(2q), R^(3q), R^(4q), and R^(5q) isindependently selected from the group consisting of H and C₁-C₆ alkyl.2. A compound of claim 1 wherein G is selected from the group consistingof: (1) —C≡C—, (2) —C≡C—C≡C—, (3) —CR^(3G)═CR^(3G)—C≡C—, and (4)—C≡C—CR^(3G)═CR^(3G)—.
 3. A compound of claim 2 wherein G is selectedfrom the group consisting of: (1) —C≡C—, (2) —C≡C—C≡C—, (3) —CH═CH—C≡C—,and (4) —C≡C—CH═CH—.
 4. A compound of claim 3 wherein G is —C≡C—.
 5. Acompound of claim 3 wherein G is —C≡C—C≡C—.
 6. A compound of claim 3wherein G is —CH═CH—C≡C—.
 7. A compound of claim 6 wherein G has thefollowing structure:


8. A compound of claim 3 wherein G is —C≡C—CH═CH—.
 9. A compound ofclaim 8 wherein G has the following structure:


10. A compound of any one of claims 1-9 wherein X is —(C═O)NR₄—.
 11. Acompound of claim 10 wherein X is —(C═O)NH—.
 12. A compound of any oneof claims 1-11 wherein Q is —(C═O)N(R₁,R₂).
 13. A compound of claim 12wherein Q is —(C═O)NHOH.
 14. A compound of any one of claims 1-13wherein n is
 0. 15. A compound of any one of claims 1-14 wherein R₃ isH.
 16. A compound of any one of claims 1-15 wherein Y is substituted orunsubstituted aryl.
 17. A compound of claim 16 wherein Y is substitutedor unsubstituted phenyl.
 18. A compound of claim 17 wherein Y isunsubstituted phenyl.
 19. A compound of any one of claims 1-18 wherein Ais selected from the group consisting of: (1)—(CH₂)₀₋₄C(R^(1a),R^(2a))(CH₂)₀₋₄OR^(3a), (2)—(CH₂)₀₋₄C(R^(1a),R^(2a))N(R^(4a),R^(5a)), and (3) —CH(R^(1a),R^(2a)).20. A compound of claim 19 wherein A is selected from the groupconsisting of —CH(CH₃)₂, —CH₂OH, —CH₂NH₂, —CHCH₃OH, —CHCH₃NH₂ and—C(CH₃)₂OH.
 21. A compound of claim 19 wherein A is —C(CH₃)₂NH₂.
 22. Acompound of any one of claims 1-18 wherein A is selected from the groupconsisting of: (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl, (2)substituted or unsubstituted aryl, (3) substituted or unsubstitutedheterocyclyl, and (4) substituted or unsubstituted heteroaryl.
 23. Acompound of claim 22 wherein A is selected from the group consisting of


24. A compound of any one of claims 1-23 wherein R₃ is H and

has the following structure:


25. A compound of claim 24 wherein A is —CHCH₃OH or —CHCH₃NH₂

and has one of the following structures:


26. A compound of any one of claims 1-25 wherein D is present.
 27. Acompound of claim 26 wherein D is substituted or unsubstitutedheteroaryl.
 28. A compound of claim 27 wherein heteroaryl is selectedfrom the group consisting of:


29. A compound of claim 26 wherein D is substituted or unsubstitutedaryl.
 30. A compound of claim 29 wherein D is substituted orunsubstituted phenyl.
 31. A compound of claim 30 wherein D isunsubstituted phenyl.
 32. A compound of any one of claims 1-25 wherein Dis absent.
 33. A compound of any one of claims 1-32 wherein L ispresent.
 34. A compound of claim 33 wherein L is substituted orunsubstituted alkyl
 35. A compound of claim 34 wherein wherein L is—CH₃—.
 36. A compound of claim 33 wherein L is —CH₂—NH—.
 37. A compoundof any one of claims 1-32 wherein L is absent.
 38. A compound having thefollowing formula (I):

or a stereoisomer, pharmaceutically acceptable salt, ester, or prodrugthereof, wherein: E is selected from the group consisting of: (1) H, (2)substituted or unsubstituted C₁-C₆-alkyl, (3) substituted orunsubstituted C₂-C₆-alkenyl, (4) substituted or unsubstitutedC₂-C₆-alkynyl, (5) substituted or unsubstituted C₃-C₁₀-cycloalkyl, (6)substituted or unsubstituted aryl, (7) substituted or unsubstitutedheterocyclyl, and (8) substituted or unsubstituted heteroaryl; L isabsent or selected from the group consisting of: (1) substituted orunsubstituted C₁-C₆-alkyl, (2) —(NR^(3L))₀₋₁—(CH₂)₀₋₄—NR^(3L)—(CH₂)₀₋₄—,(3) —(NR^(3L))₀₋₁—C(R^(1L),R^(2L))—NR^(3L)C(R^(1L),R^(2L))—, (4)—C(R^(1L),R^(2L))—O—C(R^(1L),R^(2L))—, (5) —(CH₂)₀₋₄—NR^(3L)—C(R^(1L),R^(2L))—CONH—(CH₂)₀₋₄—, (6) —CO—C(R^(1L),R^(2L))—NHCO—, (7) —CONR^(3L)—,(8) —NR^(3L)CO—, (9) —NR^(3L)—, (10) —SO₂NR^(3L)—, (11)—NR^(3L)—C(═O)—NR^(3L)—, (12) substituted or unsubstitutedC₃-C₁₀-cycloalkyl, (13) substituted or unsubstituted aryl, (14)substituted or unsubstituted heterocyclyl, and (15) substituted orunsubstituted heteroaryl, wherein: each R^(1L), R^(2L), and R^(3L) isindependently selected from the group consisting of:  (a) H,  (b)substituted or unsubstituted C₁-C₆-alkyl,  (c) C₁-C₆-alkyl substitutedwith aryl,  (d) C₁-C₆-alkyl substituted with heterocyclyl, and  (e)C₁-C₆-alkyl substituted with heteroaryl, or R^(1L) and R^(3L), togetherwith the atoms to which they are attached can form a substituted orunsubstituted heterocyclic ring, having from 3 to 8 ring atoms, wherein1-2 ring atoms of the heterocyclic ring are selected from N, O and S; Dis absent or selected from the group consisting of: (1) substituted orunsubstituted C₃-C₁₀-cycloalkyl, (2) substituted or unsubstituted aryl,(3) substituted or unsubstituted heterocyclyl, and (4) substituted orunsubstituted heteroaryl; G is selected from the group consisting of:(1) —(CH₂)₀₋₄—O—(CH₂)₀₋₄—, (2) —(CH₂)₀₋₄—S—(CH₂)₀₋₄—, (3)—(CH₂)₀₋₄—NR^(1G)—(CH₂)₀₋₄—, (4) —C(═O)—, (5) —NR^(1G)C(═O)—, (6)—C(═O)NR^(1G)—, (7) —(CH₂)₀₋₄NHCH₂C(═O)NR^(1G)—, (8) —C≡C—, (9)—C≡C—C≡C—, (10) —CR^(2G)═CR^(2G)—, (11) —S(═O)—, (12) —SO₂—, (13)—C(R^(3G))₂—S(═O)—, (14) —S(═O)—C(R^(3G))₂—, (15) —C(R^(3G))₂—SO₂—, (16)—SO₂—C(R^(3G))₂—, (17) —CR^(3G)═CR^(3G)—CR^(3G)═CR^(3G)—, (18)—C(R^(3G))₂—, (19) —CR^(3G)═CR^(3G)—C≡C—, (20) —C≡C—CR^(3G)═CR^(3G)—,(21) —C(═O)—C≡C—, (22) —C≡C—C(═O)—, (23) substituted or unsubstitutedC₃-C₁₀-cycloalkyl, (24) substituted or unsubstituted aryl, (25)substituted or unsubstituted heterocyclyl, and (26) substituted orunsubstituted heteroaryl, wherein: R^(1G) is substituted orunsubstituted C₁-C₆-alkyl; each R^(2G) and R^(3G) is independentlyselected from the group consisting of H, a halogen atom, and substitutedor unsubstituted C₁-C₆-alkyl; Y is absent or selected from the groupconsisting of: (1) substituted or unsubstituted C₃-C₁₀-cycloalkyl, (2)substituted or unsubstituted aryl, (3) substituted or unsubstitutedheterocyclyl, and (4) substituted or unsubstituted heteroaryl; X isselected from the group consisting of: (1) —(C═O)NR₄—, (2)—C₁-C₆-alkyl-(C═O)NR₄—, (3) —C₂-C₆-alkenyl-(C═O)NR₄—, (4)—C₂-C₆-alkynyl-(C═O)NR₄—, (5) —CH₂NR₄—, (6) —SO₂NR₄—, (7) —S(═O)NR₄—,(8) —NR₄C(═O)—, and (9) —NR₄—, or X and A, together with the atoms towhich they are attached can form a heterocyclic ring, having from 5 to 8ring atoms, wherein 1-2 ring atoms of the heterocyclic ring are selectedfrom N, O and S, or when Y is a bicyclic substituted or unsubstitutedheterocyclyl or heteroaryl, then X is absent; R₃ is H or substituted orunsubstituted C₁-C₆-alkyl, or R₃ and A, together with the atom to whichthey are attached can form a substituted or unsubstituted 3-10 memberedcycloalkyl or a heterocyclic ring, having from 3 to 10 ring atoms,wherein 1-4 ring atoms of the heterocyclic ring are selected from N, Oand S; R₄ is (1) H or substituted or unsubstituted C₁-C₆-alkyl, or (2)R₄ and A, together with the atoms to which they are attached can form asubstituted or unsubstituted heterocyclic ring, having from 3 to 8 ringatoms, wherein 1-2 ring atoms of the heterocyclic ring are selected fromN, O and S, or (3) R₄ and Y, together with the atoms to which they areattached, form a bicyclic substituted or unsubstituted heterocyclyl orheteroaryl; n is an integer from 0-6; A is selected from the groupconsisting of: (1) —C(R^(1a),R^(2a))OR^(3a), (2)—C(R^(1a),R^(2a))N(R^(4a),R^(5a)), (3) substituted or unsubstitutedC₃-C₁₀-cycloalkyl, (4) substituted or unsubstituted aryl, (5)substituted or unsubstituted heterocyclyl, and (6) substituted orunsubstituted heteroaryl, wherein: each R^(1a) and R^(2a) isindependently selected from the group consisting of substituted orunsubstituted C₁-C₆-alkyl; each R^(1a), R^(4a), and R^(5a) isindependently selected from the group consisting of:  (a) H,  (b) ahalogen atom,  (c) substituted or unsubstituted C₁-C₆-alkyl,  (d)substituted or unsubstituted aryl,  (e) substituted or unsubstitutedheterocyclyl, and  (f) substituted or unsubstituted heteroaryl, orR^(4a) and R^(5a) together with the N atom to which they are attachedcan form a substituted or unsubstituted heterocyclic ring, having from 5to 8 ring atoms, wherein 1-2 ring atoms of the heterocyclic ring areselected from N, O and S; and when A is —C(R^(1a),R^(2a))OR^(3a), thecompound is not2-{[(4′-ethyl-1,1′-biphenyl-4-yl)carbonyl]amino}-3-hydroxy-3-methylbutanoicacid, 4′-ethyl-N-{2-hydroxy-1-[(hydroxyamino)carbonyl]-2-methylpropyl}-1,1′-biphenyl-4-carboxamide orN-{2-hydroxy-1-[(hydroxyamino)carbonyl]-2-methylpropyl}-4-(phenylethynyl)benzamide;Q is absent or selected from the group consisting of: (1)—C(═O)N(R₁,R₂), (2) —NHC(═O)N(R₁,R₂), (3) —N(OH)C(═O)N(R₁,R₂), (4)—CH(OH)C(═O)N(R₁,R₂), (5) —CH[N(R^(2q), R^(3q))]C(═O)N(R₁,R₂), (6)—CHR^(1q)C(═O)N(R₁, R₂), (7) —CO₂H, (8) —C(═O)NHSO₂R^(4q), (9) —SO₂NH₂,(10) —N(OH)C(═O)R^(1g), (11) —N(OH)SO₂R^(4q), (12) —NHSO₂R^(4q), (13)—SH, (14) —CH(SH)(CH₂)₀₋₄C(═O)N(R₁,R₂), (15) —CH(SH)(CH₂)₀₋₁CO₂H, (16)—CH(OH)(CH₂)₀₋₁CO₂H, (17) —CH(SH)CH₂CO₂R^(1q), (18) —CH(OH)(CH₂)SO₂NH₂,(19) —CH(CH₂SH)NHCOR^(1q), (20) —CH(CH₂SH)NHSO₂R^(4q), (21)—CH(CH₂SR^(5q))CO₂H, (22) —CH(CH₂SH)NHSO₂NH₂, (23) —CH(CH₂OH)CO₂H, (24)—CH(CH₂OH)NHSO₂NH₂, (25) —C(═O)CH₂CO₂H, (26) —C(═O)(CH₂)₀₋₁CONH₂, (27)—OSO₂NHR^(5q), (28) —SO₂NHNH₂, (29) —P(═O)(OH)₂,

and (33) —N(OH)C(═O)CR₁R₂, wherein: R₁ is selected from the groupconsisting of:  (1) —H,  (2) —OH,  (3) —OC₁-C₆-alkyl,  (4) —N(R^(2q),R^(3q)), and  (5) substituted or unsubstituted C₁-C₆-alkyl; R₂ isselected from the group consisting of:  (1) H,  (2) substituted orunsubstituted C₁-C₆-alkyl,  (3) substituted or unsubstitutedC₂-C₆-alkenyl,  (4) substituted or unsubstituted C₂-C₆-alkenyl,  (5)substituted or unsubstituted aryl,  (6) substituted or unsubstitutedheterocyclyl, and  (7) substituted or unsubstituted heteroaryl, or R₁and R₂, together with the N atom to which they are attached can form asubstituted or unsubstituted heterocyclic ring, having from 3 to 10 ringatoms, wherein 1-4 ring atoms of the heterocyclic ring are selected fromN, O and S; and each R^(1q), R^(2q), R^(3q), R^(4q), and R^(5q) isindependently selected from the group consisting of H and C₁-C₆ alkyl.39. A compound of claim 38 wherein A is—C(R^(1a),R^(2a))N(R^(4a),R^(5a)).
 40. A compound of claim 39 wherein Ais —C(CH₃)₂NH₂.
 41. A compound of claim 38 wherein A is—C(R^(1a),R^(2a))OR^(3a).
 42. A compound of claim 38 wherein A isselected from the group consisting of: (1) substituted or unsubstitutedC₃-C₁₀-cycloalkyl, (2) substituted or unsubstituted aryl, (3)substituted or unsubstituted heterocyclyl, and (4) substituted orunsubstituted heteroaryl.
 43. A compound of claim 42 wherein A isselected from the group consisting of


44. A compound of any one of claims 38-43 wherein R₃ is H and

has the following structure:


45. A compound of any one of claims 38-44 wherein G is selected from thegroup consisting of: (1) —C≡C—, (2) —C≡C—C≡C—, (3)—CR^(3G)═CR^(3G)—C≡C—, and (4) —C≡C—CR^(3G)═CR^(3G)—.
 46. A compound ofclaim 45 wherein G is selected from the group consisting of: (1) —C≡C—,(2) —C≡C—C≡C—, (3) —CH═CH—C≡C—, and (4) —C≡C—CH═CH—.
 47. A compound ofclaim 46 wherein G is —C≡C—.
 48. A compound of claim 46 wherein G is—C≡C—C≡C—C≡C—.
 49. A compound of claim 46 wherein G is —CH═CH—C≡C—. 50.A compound of claim 49 wherein G has the following structure:


51. A compound of claim 46 wherein G is —C≡C—CH═CH—.
 52. A compound ofclaim 51 wherein G has the following structure:


53. A compound of any one of claims 38-52 wherein X is —(C═O)NR₄—.
 54. Acompound of claim 53 wherein X is —(C═O)NH—.
 55. A compound of any oneof claims 38-54 wherein Q is —(C═O)N(R₁,R₂).
 56. A compound of claim 55wherein Q is —(C═O)NHOH.
 57. A compound of any one of claims 38-56wherein n is
 0. 58. A compound of any one of claims 38-57 wherein R₃ isH.
 59. A compound of any one of claims 38-58 wherein Y is substituted orunsubstituted aryl.
 60. A compound of claim 59 wherein Y is substitutedor unsubstituted phenyl.
 61. A compound of claim 60 wherein Y isunsubstituted phenyl.
 62. A compound of any one of claims 38-61 whereinD is present.
 63. A compound of claim 62 wherein D is substituted orunsubstituted heteroaryl.
 64. A compound of claim 63 wherein heteroarylis selected from the group consisting of:


65. A compound of claim 62 wherein D is substituted or unsubstitutedaryl.
 66. A compound of claim 65 wherein D is substituted orunsubstituted phenyl.
 67. A compound of claim 66 wherein D isunsubstituted phenyl.
 68. A compound of any one of claims 38-61 whereinD is absent.
 69. A compound of any one of claims 38-68 wherein L ispresent.
 70. A compound of claim 69 wherein L is substituted orunsubstituted alkyl
 71. A compound of claim 70 wherein wherein L is—CH₃—.
 72. A compound of claim 69 wherein L is —CH₂—NH—.
 73. A compoundof any one of claims 38-68 wherein L is absent.
 74. A pharmaceuticalcomposition comprising a compound of any one of claims 1-73 and apharmaceutically acceptable carrier or diluent.
 75. A method fortreating a subject with a gram-negative bacterial infection comprisingadministering to the subject in need thereof an antibacteriallyeffective amount of a compound of any one of claims 1-73 or apharmaceutical composition of claim
 74. 76. A method for treating asubject with a gram-negative bacterial infection comprisingco-administering to the subject in need thereof an antibacteriallyeffective amount of a compound of any one of claims 1-73 or apharmaceutical composition of claim 74 and a second antibacterial agent.